Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) is a preadsorbed secondary antibody with a maximum absorption wavelength of 590nm and a maximum emission wavelength of 617nm. Ideal for fluorescent cell and tissue imaging. Suitable for IHC, ICC/IF, Flow Cytometry and ELISA.
- Preadsorbed to prevent cross-reactivity and reduce background staining
- Intense fluorescence and high photostability allowing more time for image capture especially when detecting low abundance targets
- Minimal spectral overlap with other Alexa Fluor® dyes makes this ideal for use in multi-color analysis
- Cited in over 100 publications
Application | Reactivity | Dilution info | Notes |
---|---|---|---|
Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
Application IHC-Fr | Reactivity Reacts | Dilution info - | Notes - |
Application IHC-P | Reactivity Reacts | Dilution info - | Notes - |
Application ICC/IF | Reactivity Reacts | Dilution info 1/200.00000 - 1/1000.00000 | Notes - |
Application Flow Cyt | Reactivity Reacts | Dilution info 1/2000 | Notes ab178000 - Mouse monoclonal IgG1 (Alexa Fluor® 594), is suitable for use as an isotype control to complement this secondary antibody. |
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Igh-4, Ighg1, Ig gamma-1 chain C region secreted form
Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) is a preadsorbed secondary antibody with a maximum absorption wavelength of 590nm and a maximum emission wavelength of 617nm. Ideal for fluorescent cell and tissue imaging. Suitable for IHC, ICC/IF, Flow Cytometry and ELISA.
- Preadsorbed to prevent cross-reactivity and reduce background staining
- Intense fluorescence and high photostability allowing more time for image capture especially when detecting low abundance targets
- Minimal spectral overlap with other Alexa Fluor® dyes makes this ideal for use in multi-color analysis
- Cited in over 100 publications
By immunoelectrophoresis and ELISA this antibody reacts specifically with mouse IgG and with light chains common to other mouse immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. Reduced cross-reactivity to bovine, chicken, horse, human, pig, rabbit and rat IgG was detected. This antibody may cross react with IgG from other species.
Preservative: 0.02% Sodium azide
Constituents: PBS, 23% Glycerol (glycerin, glycerine), 1% BSA
Antiserum was cross adsorbed using bovine, chicken, horse, human, pig, rabbit and rat immunosorbents to remove cross reactive antibodies. The antibody to mouse IgG was isolated by affinity chromatography using antigen coupled to agarose beads.
This antibody reacts with the heavy and light chains of Mouse IgG
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Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.
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BrdU immunofluorescence staining of HeLa cells using rat anti-BrdU antibody
Anti-BrdU antibody [BU1/75 (ICR1)] - Proliferation Marker ab6326 staining BrdU in Hela cells. Untreated and BrdU treated (10uM for 24 hours) cells. The cells were fixed with 100% methanol (5 min) and then subjected to acid hydrolysis using 2M HCL in 0.1% PBS-Tween for 30 minutes at room temperature to denature the DNA. They were then permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1hr. The cells were then incubated overnight at 4°C with Anti-BrdU antibody [BU1/75 (ICR1)] - Proliferation Marker ab6326 at 1μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preadsorbed ab150165 Goat polyclonal Secondary Antibody to Rat IgG - H&L (Alexa Fluor® 488) pre-adsorbed at 1/1000 dilution (shown in green) and ab150120 Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling Estrogen Receptor alpha with purified Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade ab32063 at 1/1000. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. The cells were co-stained with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) using ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) as the secondary antibody. Nuclei counterstained with DAPI (blue).
Control 1: primary antibody (1/1000) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).
ICC/IF image of Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, 1μg/ml) overnight at +4°C. The secondary antibody (orange) was ab150120 Alexa Fluor® 594 goat anti-mouse IgG (H+L) used at 1μg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.
Immunocytochemistry/Immunofluorescence analysis of A431 (human epidermoid carcinoma) cells labeling alpha smooth muscle Actin (green) with purified Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] ab32575 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counterstained with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, anti-Tubulin (mouse mAb) at 1/1000 followed by ab150120 Alexa Fluor®594 goat anti-mouse secondary (1/1000). Nuclei were counterstained with DAPI (blue).
For negative control 1, rabbit primary antibody and anti-mouse secondary antibody (ab150120) were used. For negative control 2, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (mouse primary antibody) was used followed by anti-rabbit secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077).
Image: Courtesy of Dr. Shaohua Li, UMDNJ-Robert Wood Johnson Medical School
Sample: mouse embryonic stem cell-differentiated embryoid bodies (EBs)
Preparation:
Fix in 3%PFA in PBS for 30 min at RTIncubate in 7.5% sucrose-PBS for 3h at RTIncubate in 15% sucrose-PBS at 4 degree Celsius overnightEmbed the EBs in tissue-Tek OCT compoundCut frozen sections to 4-20 μm thickness
Primary antibody 1: Rabbit anti-laminin, 1:400
Primary antibody 2: Mouse anti-disabled-2, 1:100
Secondary antibody 1: Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) pre-adsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081), 1:200
Secondary antibody 2: Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) pre-adsorbed (ab150120), 1:200
Nuclei were counterstained with DAPI
Immunofluorescence staining of GABA Transporter 1 / GAT 1 using Anti-GABA Transporter 1 / GAT 1 antibody [EPR24202-20] ab259971 in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h.
The cells were then incubated overnight at +4°C with Anti-GABA Transporter 1 / GAT 1 antibody [EPR24202-20] ab259971 at 1 μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.
Immunofluorescence staining of GABA Transporter 1 / GAT 1 using Anti-GABA Transporter 1 / GAT 1 antibody [EPR24202-20] ab259971 in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h.
The cells were then incubated overnight at +4°C with Anti-GABA Transporter 1 / GAT 1 antibody [EPR24202-20] ab259971 at 5 μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Mouse splenocyte cells labelling CD18 with Anti-CD18 antibody [EPR29884-566] ab324101 at 1/50 (10.18 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).
Confocal image showing cytoplasmic staining in a subset of mouse splenocytes (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
-ve control 1: Anti-CD18 antibody [EPR29884-566] ab324101 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.
Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling TRPML1/MG-2 with Anti-TRPML1/MG-2 antibody [EPR28651-127] ab323642 at 1/100 (5.07 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 1000 2ug/ml dilution (Green).
Confocal image showing lysosomal staining in Neuro-2a cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Low expression: F9
Anti-LAMP2 antibody [H4B4] - Lysosome Marker ab25631 Anti-LAMP2 mouse monoclonal antibody - Lysosome Marker was used to counterstain tubulin at 1/50 2ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/ 1000 2ug/ml dilution.
-ve control 1: Anti-TRPML1/MG-2 antibody [EPR28651-127] ab323642 at 1/100 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2: Anti-LAMP2 antibody [H4B4] - Lysosome Marker ab25631 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage)) cells labelling CD18 with Anti-CD18 antibody [EPR29884-566] ab324101 at 1/50 (10.18 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).
Confocal image showing cytoplasmic staining in RAW 264.7 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression: Neuro-2a.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
-ve control 1: Anti-CD18 antibody [EPR29884-566] ab324101 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neural/glia cell cells labelling Synaptojanin with Anti-Synaptojanin antibody [EPR28576-183] ab323848 at 1/500 (1.062 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 1000 2ug/ml dilution (Green).
Confocal image showing cytoplasmic staining in rat primary neural/glia cell (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Confocal scanning Z step was set as 0.3 um followed by image processing with maximum Z projection. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 500 4ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 1000 2ug/ml dilution.
-ve control 1: Anti-Synaptojanin antibody [EPR28576-183] ab323848 at 1/500 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2: Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling Synaptojanin with Anti-Synaptojanin antibody [EPR28576-183] ab323848 at 1/500 (1.062 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 1000 2ug/ml dilution (Green).
Confocal image showing cytoplasmic staining in mouse primary neural/glia cell (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Confocal scanning Z step was set as 0.3 um followed by image processing with maximum Z projection.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 500 4ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 1000 2ug/ml dilution.
-ve control 1: Anti-Synaptojanin antibody [EPR28576-183] ab323848 at 1/500 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2: Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neural/glia cells labelling EAAT2 with Anti-EAAT2 antibody [EPR29552-528] ab323867 at 1/100 dilution (5.26 ug/ml), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/ml) (Green).
Confocal image showing positive staining in rat primary glia cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
ab10062 Anti-GFAP mouse monoclonal antibody was used to counterstain tubulin at 1/50 dilution (10 ug/ml), followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (2 ug/ml) (Magenta).
-ve control 1: Anti-EAAT2 antibody [EPR29552-528] ab323867 at a 1/100 dilution followed by ab150120 at a 1/1000 dilution.
-ve control 2: ab10062 at a 1/50 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 at a 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SK-OV-3(human ovarian cancer epithelial cell) cells labelling PINK1 with Anti-PINK1 antibody [EPR29146-340] ab323807 at 1/50 (10.34 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 g/ml dilution (Green).
Confocal image showing mitochondrial staining in SK-OV-3 cells (shown in green) treatment with 30 μM CCCP for 6 hours. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 g/ml dilution.
anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/200 2.5 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Counterstain Secondary antibody only control: Secondary antibody is ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 2 g/ml dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling VAPB with Anti-VAPB antibody [EPR27026-59] ab324047 at 1/50 (9.76 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).
Confocal image showing cytoplasmic staining in HeLa cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
Anti-KDEL mouse monoclonal antibody was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 (2 ug/ml) dilution (Magenta).
-ve control 1: Anti-VAPB antibody [EPR27026-59] ab324047 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2: Anti-KDEL mouse monoclonal antibody at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse cardiomyocyte cells labelling Natriuretic peptides A with Anti-Natriuretic peptides A antibody [EPR22089-283] ab225844 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor®:488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibody at 1/1000 dilution.
Confocal image showing cytoplasmic staining in a subset of mouse cardiomyocytes (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-Sarcomeric Alpha Actinin antibody [EA-53] ab9465 Anti-Sarcomeric Alpha Actinin mouse monoclonal antibody was used to counterstain Sarcomeric Alpha Actinin at 1/200.
The negative controls are as follows:
-ve control 1: Anti-Natriuretic peptides A antibody [EPR22089-283] ab225844 at 1/100 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) secondary antibody name (ab150120) secondary at 1/1000 dilution.
-ve control 2: Primary (Anti-Sarcomeric Alpha Actinin antibody [EA-53] ab9465) at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary at 1/200 dilution.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Human tonsil (fresh) tissue labeling FOXP3 with Anti-FOXP3 antibody [mAbcam 450] - Mouse IgG2a ab309108 at 1/10000 dilution followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 2 ug/mL dilution (Green). Confocal image showing positive staining on human tonsil. The nuclear counterstain was DAPI (Blue). The section was incubated with Anti-FOXP3 antibody [mAbcam 450] - Mouse IgG2a ab309108 overnight at 4°C. The section was then mounted using Fluoromount®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control: Secondary antibody is ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594)at 1/1000 2 ug/mL dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse splenocyte cells labelling Neogenin with Anti-Neogenin antibody [EPR30444-522] ab324107 at 1/50 (9.76 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).
Low expression: confocal image showing no staining in mouse splenocytes (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
-ve control 1: Anti-Neogenin antibody [EPR30444-522] ab324107 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling Cytochrome C oxidase subunit II with Anti-MTCO2 antibody [EPR3313] ab109739 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibody at 1/1000 dilution.
Confocal image showing mitochondria staining in HepG2 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain COX IV at 1/1000 dilution (Magenta).
The negative controls are as follows:
-ve control 1: Anti-MTCO2 antibody [EPR3313] ab109739 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary at 1/1000 dilution.
-ve control 2: anti-COX IV mouse monoclonal antibody - Mitochondrial Marker antibody at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary at 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat cardiomyocyte cells labelling Natriuretic peptides A with Anti-Natriuretic peptides A antibody [EPR22089-283] ab225844 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor®:488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibody at 1/1000 dilution.
Confocal image showing cytoplasmic staining in a subset of rat cardiomyocytes (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-Sarcomeric Alpha Actinin antibody [EA-53] ab9465 Anti-Sarcomeric Alpha Actinin mouse monoclonal antibody was used to counterstain Sarcomeric Alpha Actinin at 1/200.
The negative controls are as follows:
-ve control 1: Anti-Natriuretic peptides A antibody [EPR22089-283] ab225844 at 1/100 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary at 1/1000 dilution.
-ve control 2: Anti-Sarcomeric Alpha Actinin antibody [EA-53] ab9465 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary at 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized T-47D (human ductal breast epithelial tumor epithelial cell) cells labelling Quiescin Q6 with Anti-Quiescin Q6 antibody [EPR25653-56] ab323533 at 1/50 (9.95 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).
Confocal image showing cytoplasmic staining in T-47D cell line (shown in green), which is co-localised with golgi marker. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-GM130 antibody - BSA and Azide free ab169276 Anti-GM130 mouse polyclonal antibody - cis-Golgi Marker was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
-ve control 1: Anti-Quiescin Q6 antibody [EPR25653-56] ab323533 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2: Anti-GM130 antibody - BSA and Azide free ab169276 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neural/glia cell cells labelling Contactin 1 with ab323505 at 1/50 (10.36 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).
Confocal image showing cytoplasmic staining in rat primary neuron (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
-ve control 1: ab323505 at 1/200 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2: Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MCF7 (human breast adenocarcinoma epithelial cell) cells labelling Quiescin Q6 with Anti-Quiescin Q6 antibody [EPR25653-56] ab323533 at 1/50 (9.95 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).
Confocal image showing cytoplasmic staining in MCF7 cell line (shown in green), which is co-localised with golgi marker. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-GM130 antibody - BSA and Azide free ab169276 Anti-GM130 mouse polyclonal antibody - cis-Golgi Marker was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
-ve control 1: Anti-Quiescin Q6 antibody [EPR25653-56] ab323533 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2: Anti-GM130 antibody - BSA and Azide free ab169276 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Human PBMC (human peripheral blood mononuclear cell) cells labelling Quiescin Q6 with Anti-Quiescin Q6 antibody [EPR25653-56] ab323533 at 1/50 (9.95 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).
Negative control: Confocal image showing no staining in human PBMCs (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-GM130 antibody - BSA and Azide free ab169276 Anti-GM130 mouse polyclonal antibody - cis-Golgi Marker was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
-ve control 1: Anti-Quiescin Q6 antibody [EPR25653-56] ab323533 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2: Anti-GM130 antibody - BSA and Azide free ab169276 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) cells labelling Quiescin Q6 with Anti-Quiescin Q6 antibody [EPR25653-56] ab323533 at 1/50 (9.95 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).
Confocal image showing cytoplasmic staining in PANC-1 cell line and no staining in HEK-293 cell line (shown in green), which is co-localised with golgi marker. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-GM130 antibody - BSA and Azide free ab169276 Anti-GM130 mouse polyclonal antibody - cis-Golgi Marker was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
-ve control 1: Anti-Quiescin Q6 antibody [EPR25653-56] ab323533 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2: Anti-GM130 antibody - BSA and Azide free ab169276 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling Contactin 1 with ab323505 at 1/50 (10.36 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).
Confocal image showing cytoplasmic staining in mouse primary neuron (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
-ve control 1: ab323505 at 1/200 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2: Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling TRIM32 with Anti-TRIM32 antibody [EPR29683-521] ab322979 at 1/50 (10.14 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).
Confocal image showing cytoplasmic staining in mouse primary neuron (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4ug/ml dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 2ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
-ve control 1: Anti-TRIM32 antibody [EPR29683-521] ab322979 at a 1/50 dilution followed by ab150120 at a 1/1000 dilution.
-ve control 2: Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 at a 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 at a 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neuron cells labelling KIF5A with Anti-KIF5A antibody [EPR29540-31] ab322900 at 1/50 (10.14 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).
Confocal image showing cytoplasmic staining in rat primary neuron (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4ug/ml dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 2ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
-ve control 1: Anti-KIF5A antibody [EPR29540-31] ab322900 at a 1/50 dilution followed by ab150120 at a 1/1000 dilution.
-ve control 2: Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 at a 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 at a 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling KIF5A with Anti-KIF5A antibody [EPR29540-31] ab322900 at 1/50 (10.14 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).
Confocal image showing cytoplasmic staining in mouse primary neuron (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4ug/ml dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 2ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
-ve control 1: Anti-KIF5A antibody [EPR29540-31] ab322900 at a 1/50 dilution followed by ab150120 at a 1/1000 dilution.
-ve control 2: Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 at a 1/500 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 at a 1/1000 dilution.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cells labelling SCRN1 with Anti-SCRN1 antibody [EPR29587-538] ab323711 at 1/100 (4.98 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).
Confocal image showing cytoplasmic staining in mouse primary neural/glia cell (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Confocal scanning Z step was set as 0.3 ?m followed by image processing with maximum Z projection. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
-ve control 1: Anti-SCRN1 antibody [EPR29587-538] ab323711 at 1/100 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody preadsorbed at 1/1000 2 ug/ml dilution
-ve control 2: Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4ug/ml dilution , followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed.
Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] ab288063 staining ERK1 (phospho T202 + Y204) + ERK2 (phospho T185 + Y187) in untreated, PMA treated and PMA + LP treated NIH/3T3 cells. The cells were fixed with 100% Methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] ab288063 at 5µg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
The image shows increased nuclear staining after 24hr serum starvation followed by treatment with PMA (200nM, 15min) of NIH/3T3 cells. The Lambda Phoshatase treatment then removes all staining of antibody with no phospho ERK1/2 remaining.
Anti-ERK1 + ERK2 antibody [EPR17526] ab184699 was used as a Pan ERK1 + ERK2 control for Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] ab288063. The results showed nuclear staining on untreated, with no increase after PMA treatment and no reduction after PMA + LP treated in NIH/3T3 cells.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neural/glia cells labelling SCRN1 with Anti-SCRN1 antibody [EPR29587-538] ab323711 at 1/100 (4.98 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).
Confocal image showing cytoplasmic staining in rat primary neural/glia (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Confocal scanning Z step was set as 0.3 um followed by image processing with maximum Z projection. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
-ve control 1: Anti-SCRN1 antibody [EPR29587-538] ab323711 at 1/100 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 2 ug/ml dilution
-ve control 2: Anti-MAP2 antibody [HM-2] - Neuronal Marker ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4ug/ml dilution , followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Human heart (fresh) tissue labeling FOXP3 with Anti-FOXP3 antibody [mAbcam 450] - Mouse IgG2a ab309108 at 1/10000 dilution followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 2 ug/mL dilution (Green). Negative control: confocal image showing no staining on human heart. The nuclear counterstain was DAPI (Blue). The section was incubated with Anti-FOXP3 antibody [mAbcam 450] - Mouse IgG2a ab309108 overnight at 4°C. The section was then mounted using Fluoromount®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control: Secondary antibody is ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594)at 1/1000 2 ug/mL dilution.
Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] ab288063 staining ERK1 (phospho T202 + Y204) + ERK2 (phospho T185 + Y187) in untreated, PMA treated and PMA + LP treated A-10 cells. The cells were fixed with 100% Methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] ab288063 at 5µg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
The image shows increased nuclear staining after 24hr serum starvation followed by treatment with PMA (200nM, 15min) of A-10 cells. The Lambda Phoshatase treatment then removes all staining of antibody with no phospho ERK1/2 remaining.
Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] ab288063 staining ERK1 (phospho T202 + Y204) + ERK2 (phospho T185 + Y187) in untreated, PMA treated and PMA + LP treated HeLa cells. The cells were fixed with 100% Methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] ab288063 at 5µg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
The image shows increased nuclear staining after 24hr serum starvation followed by treatment with PMA (200nM, 15min) of HeLa cells. The Lambda Phoshatase treatment then removes all staining of antibody with no phospho ERK1/2 remaining.
Anti-ERK1 + ERK2 antibody [EPR17526] ab184699 was used as a Pan ERK1 + ERK2 control for Anti-ERK1 (phospho T202+Y204) + ERK2 (phospho T185+Y187) antibody [EPR25015-F2] ab288063. The results showed nuclear staining on untreated, with no increase after PMA treatment and no reduction after PMA + LP treated in HeLa cells.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling Neuropeptide Y with Anti-Neuropeptide Y antibody [EPR23483-18] - BSA and Azide free (Capture) ab281152 at 1/2000 (0.5 µg/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 µg/ml) dilution (Green).
Confocal image showing cytoplasmic staining in SH-SY5Y cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Negative control: 293T.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 (1.0 µg/ml) dilution followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at a 1/1000 (2 µg/ml) dilution (Magenta).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/ml) dilution.
Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 staining of Myc proto-oncogene protein in a HCT 116 cell spheroid. The cells were fixed with 4% paraformaldehyde (10 min), permeabilised with 2% Triton X-100 for 1h and then blocked with 1% BSA/10% normal goat serum/0.3M glycine 0.1% PBS-Tween overnight at room temperature. The spheroids were then incubated overnight at room temperature with Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 at 2 μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 2 μg/ml. DAPI was used as nuclear counterstain (shown in blue). As secondary antibodies Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat anti-Rabbit (Alexa Fluor® 488) (shown in green) and ab150120 Goat anti-Mouse (Alexa Fluor® 594) (shown in magenta) were used, incubated overnight at room temperature. All permeabilization, blocking and antibody incubation steps were performed using a rotary shaker.
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Similar results were observed using 100% Methanol (5min).
Anti-Ras antibody [EP1125Y] ab52939 staining of GTPase KRas in a HCT 116 cell spheroid. The cells were fixed with 4% paraformaldehyde (10 min), permeabilised with 2% Triton X-100 for 1h and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween overnight at room temperature. The spheroids were then incubated overnight at room temperature with Anti-Ras antibody [EP1125Y] ab52939 at 2 μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 2 μg/ml. DAPI was used as nuclear counterstain (shown in blue). As secondary antibodies Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat anti-Rabbit (Alexa Fluor® 488) (shown in green) and ab150120 Goat anti-Mouse (Alexa Fluor® 594) (shown in magenta) were used, incubated overnight at room temperature. All permeabilization, blocking and antibody incubation steps were performed using a rotary shaker.
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Similar results were observed using 100% Methanol (5min).
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RHOT2 KO HeLa (RHOT2 knockout human cervical adenocarcinoma epithelial cell), Human RHOT2 (MIRO2) knockout HeLa cell line ab265801 cells labelling MIRO2 with Anti-MIRO2 antibody [RM2077] ab322962 at 1/500 (0.906 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).
Confocal image showing mitochondrial staining in wildtype HeLa cells (shown in green), showing no staining in RHOT2 knockout HeLa cells. The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/200 2.5 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
-ve control 1: Anti-MIRO2 antibody [RM2077] ab322962 at 1/500 dilution, followed byab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.
-ve control 2: anti-COX IV mouse monoclonal antibody - Mitochondrial Marker at 1/200 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
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