Suitable for ELISA, IHC-Fr, IHC-P, Flow Cyt, ICC/IF. Ideal for fluorescent cell and tissue imaging. Cited in 1265 publications.
Goat
Mouse
IgG
Heavy & Light chains
Alexa Fluor® 488
Ex: 495nm, Em: 519nm
ELISA, IHC-Fr, IHC-P, Flow Cyt, ICC/IF
Polyclonal
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 Flow Cyt | Reactivity Reacts | Dilution info 1/2000 | Notes ab170190 - Mouse monoclonal IgG1 (Alexa Fluor® 488), is suitable for use as an isotype control to complement this secondary antibody. |
Application ICC/IF | Reactivity Reacts | Dilution info 1/200.00000 - 1/1000.00000 | Notes - |
Select an associated product type
Ig gamma-1 chain C region, membrane-bound form
Ig gamma-1 chain C region secreted form, Ighg1, Igh-4
Suitable for ELISA, IHC-Fr, IHC-P, Flow Cyt, ICC/IF. Ideal for fluorescent cell and tissue imaging. Cited in 1265 publications.
Ig gamma-1 chain C region secreted form, Ighg1, Igh-4
Goat Anti-Mouse IgG H&L (Alexa Fluor® 488)
Goat
Mouse
IgG
Heavy & Light chains
Alexa Fluor® 488
Ex: 495nm, Em: 519nm
ELISA, IHC-Fr, IHC-P, Flow Cyt, ICC/IF
Polyclonal
No
IgG
Liquid
Preservative: 0.02% Sodium azide
Constituents: PBS, 23% Glycerol (glycerin, glycerine), 1% BSA
Affinity purification Immunogen
This antibody was isolated by affinity chromatography using antigen coupled to agarose beads.
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle, Stable for 12 months at -20°C, Store in the dark
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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Terms & Conditions.
alpha smooth muscle Actin flow cytometry staining of SV40LT-SMC cells using mouse anti-alpha smooth muscle Actin antibody
Overlay histogram showing SV40LT-SMC cells stained with Anti-alpha smooth muscle Actin antibody [1A4] ab7817 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (Anti-alpha smooth muscle Actin antibody [1A4] ab7817 1.137μg/ml) for 30 min at 22°C. The secondary antibody used was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113) at 1/2000 dilution for 30 min at 22°C
Isotype control antibody (black line) was mouse IgG2a [18C8BC7AD10] (Mouse IgG2c, kappa monoclonal [18C8BC7AD10] - Isotype Control ab170191) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.
This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.
Overlay histogram showing HeLa cells stained with Anti-PCNA antibody [PC10] ab29 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween 20 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (Anti-PCNA antibody [PC10] ab29, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (Mouse IgG2a, Kappa Monoclonal [B12/8] - Isotype Control ab91361, 0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Overlay histogram showing HeLa cells stained with Anti-p53 antibody [PAb 240] ab26 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (Anti-p53 antibody [PAb 240] ab26, 1μg/1x10^6 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control ab91353, 2μg/1x10^6 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
ICC/IF image of Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 stained HeLa cells. The cells were 100% methanol fixed (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to 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 (green) was ab150113 Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at 2μg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43μM.
The negative control (inset) is a secondary-only assay to demonstrate low non-specific binding of the secondary antibody.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 cells labelling Mouse IgG1 with Mouse IgG1 monoclonal [R312-MouseIgG1]-Isotype control ab280974 at 1/20 dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Anti-beta Tubulin antibody [EPR16774] ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).
Confocal image showing no staining in RAW 264.7 cells.
Negative control 1: Mouse IgG1 monoclonal [R312-MouseIgG1]-Isotype control ab280974 at a 1/20 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080 at a 1/1000 dilution.
Negative control 2: Anti-beta Tubulin antibody [EPR16774] ab179513 at a 1/200 dilution followed by ab150113 at a 1/1000 dilution.
Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized Calu-3 (human lung adenocarcinoma epithelial cell) labelling (A) MUC5B with Anti-MUC5B antibody [EPR27200-159] ab315330 at 1/500 (1.008 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 (2 ug/ml) dilution (Green), or (B) MUC5AC with Anti-Mucin 5AC antibody [45M1] ab3649 at 1/100 (10.45 ug/ml) followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).
This antibody does not cross-react with human MUC5AC.
(A) No staining of MUC5B in Calu-3 cell line.
(B) Positive staining of MUC5AC (Anti-Mucin 5AC antibody [45M1] ab3649, green) in Calu-3 cell line.
Images were taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/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 (2 ug/ml) dilution.
Flow cytometric analysis of mouse peripheral blood mononuclear cells (PBMCs) cells labelling CD32B with Anti-CD32B antibody [AT130-5] ab290737 at 1/1000 dilution (0.1 μg)/ Right compared with a mouse monoclonal IgG / Left isotype control. Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/5000 dilution was used as the secondary antibody. Cells were stained with mouse IgG or Anti-CD32B antibody [AT130-5] ab290737, then stained with rabbit anti-mouse CD19 (Anti-CD19 antibody [EPR23174-145] - BSA and Azide free ab267394) conjugated to APC. The conjugation was performed by using APC Conjugation Kit - Lightning-Link® (APC Conjugation Kit - Lightning-Link® ab201807). Gated on viable cells.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse spleen (fresh) tissue labeling CD32B with Anti-CD32B antibody [AT130-5] ab290737 at 1/100 (11.1 μg/ml) dilution followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2 μg/ml) (Green). Positive staining on mouse spleen is observed. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: PBS was used instead of primary antibody followed by secondary antibody ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488)at 1/1000 dilution (2 μg/ml).
Flow cytometric analysis of Human PBMC (peripheral blood mononuclear cell) cells labelling Integrin alpha 4/CD49D with Anti-Integrin alpha 4/CD49D antibody [P4C2] ab309548 at 1/1000 dilution (0.1ug)/ Right compared with a Mouse monoclonal IgG / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells. Then stained with anti-CD3 conjugated to Alexa Fluor® 647.
Flow cytometric analysis of Integrin alpha 4/CD49D KO HAP1 (Integrin alpha 4/CD49D knockout human chronic myelogenous leukemia near-haploid cell) / parental HAP1 cell cells labelling Integrin alpha 4/CD49D with Anti-Integrin alpha 4/CD49D antibody [P4C2] ab309548 at 1/1000 dilution (0.1ug) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells.
Flow cytometric analysis of K562 (human chronic myelogenous leukemia lymphoblast, Left) / MOLT-4 (human lymphoblastic leukemia T lymphoblast, Right) cells labelling Integrin alpha 4/CD49D with Anti-Integrin alpha 4/CD49D antibody [P4C2] ab309548 at 1/1000 dilution (0.1ug) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells.
Negative control: K-562 (PMID:16105875).
Flow cytometric analysis of Raji (human Burkitt's lymphoma B lymphocyte, Left) / Jurkat (human T cell leukemia T lymphocyte from peripheral blood, Right) cells labelling CD3 epsilon with Anti-CD3 epsilon antibody [BC3] ab307971 at 1/1000 dilution (0.1 ug)/Red compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/5000 dilution was used as the secondary antibody. Negative control: Raji. Gated on viable cells.
Flow cytometric analysis of human PBMC (human peripheral blood mononuclear cell) cells labelling CD3 epsilon with Anti-CD3 epsilon antibody [BC3] ab307971 at 1/1000 dilution (0.1 ug)/Red compared with a Mouse monoclonal IgG / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/5000 dilution was used as the secondary antibody.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized LNCaP (human prostate carcinoma epithelial cell) cells labelling TMPRSS2 with Anti-TMPRSS2 antibody [P5H9-A3] ab309546 at 1/100 (10.28 ug/ml) dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in LNCaP and negative staining in 293T (PMID: 33493182).Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). Alexa Fluor® 594 Anti-beta Tubulin antibody [EPR16774] ab206369 Anti-beta Tubulin rabbit monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
antibody only control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Jurkat (human T cell leukemia T lymphocyte, Left) / Ramos (human Burkitt's lymphoma B lymphocyte, Right) cells labelling Bcl6 with Anti-Bcl6 antibody [K112-91] ab307681 at 1/1000 dilution (0.1ug) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.
Negative control: Jurkat (PMID: 8652841, 10757802).
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized fresh frozen rat cerebrum tissue labeling GFAP with Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) ab279290 at 1/200 (10 µg/ml) dilution followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 (2μg/mL) dilution. Positive staining on rat cerebrum is observed. The nuclear counterstain was DAPI (blue).
Secondary antibody control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized fresh frozen mouse cerebrum tissue labeling GFAP with Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) ab279290 at 1/200 (10 µg/ml) dilution followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 (2μg/mL) dilution. Positive staining on mouse cerebrum is observed. The nuclear counterstain was DAPI (blue).
Secondary antibody control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
Flow cytometric (Intracellular) overlay histogram showing 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling Ubiquitin with Anti-Ubiquitin antibody [P4D1] ab303664 at 1/800 dilution (0.1μg) (red line). Isotype control antibody (black line) was a mouse monoclonal IgG used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control.
Flow cytometric (Intracellular) overlay histogram showing 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labeling Ubiquitin with Anti-Ubiquitin antibody [P4D1] ab303664 at 1/800 dilution (0.1μg) (red line). Isotype control antibody (black line) was a mouse monoclonal IgG used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control.
Flow cytometric analysis of HeLa (Human cervix adenocarcinoma epithelial cell, Left) / HUVEC (Human umbilical vein endothelial cell, Right) cells labelling Integrin beta 3 with Anti-Integrin beta 3 antibody [Y2/51] ab9509 at 1/1000 dilution (0.1 ug)/Red compared with a Mouse monoclonal IgG / Black isotype control and an unlabelled control (Cell without incubation with primary antibody and secondary antibody / Blue).
Secondary antibody was Goat anti mouse IgG (Alexa Fluor® 488, ab150113) used at 1/2000 dilution.
Negative control: HeLa.
Gated on viable cells.
Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) cells labelling Integrin beta 3 with Anti-Integrin beta 3 antibody [Y2/51] ab9509 at 1/1000 dilution (0.1 ug) / Right compared with a Mouse monoclonal IgG / Left.
Goat anti mouse IgG (Alexa Fluor® 488, ab150113) was used as the secondary antibody at 1/2000 dilution.
Cells were stained with mouse IgG or Anti-Integrin beta 3 antibody [Y2/51] ab9509. Then stained with anti-CD41 conjugated to APC.
Gated on viable cells.
Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) cells labelling Human IgG with Anti-Human IgG antibody [IG266] ab200699 at 1/800 dilution (0.1 ug) / Right compared with a Mouse monoclonal IgG / Left.
Goat anti mouse IgG (Alexa Fluor® 488, ab150113) was used as the secondary antibody at 1/2000 dilution.
Cells were stained with mouse IgG or Anti-Human IgG antibody [IG266] ab200699. Then stained with anti-CD19 conjugated to Alexa Fluor® 647.
Gated on viable cells.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat spleen (fresh) tissue labeling CD2 with Anti-CD2 antibody [OX34] ab243837 at 1/100 (10.43 ug/ml) dilution followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 (2 ug/mL) dilution (Green).
Confocal image showing positive staining on rat spleen. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 (2 ug/mL) dilution
Flow cytometric analysis of Rat splenocytes cells labelling CD2 with Anti-CD2 antibody [OX34] ab243837 at 1/1000 dilution (0.1 ug)/Right compared with a Mouse monoclonal IgG / Left isotype control.
Goat anti mouse IgG (Alexa Fluor® 488, ab150113) was used as a secondary antibody at 1/2000 dilution. Gated on viable cell.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse pituitary (fresh frozen) tissue labeling ACTH with Anti-ACTH antibody [AH26] ab233953 at 1/100 (11.64 µg/ml) dilution followed by ab150113, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green).
Positive staining on mouse pituitary. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluorr® 488) at 1/1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20)
Flow cytometric analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling Histone H2B with Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade ab52484 at 1/1000 dilution (0.1µg) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.
Flow cytometric analysis of NIH/3T3 (mouse embryonic fibroblast) cells labelling Histone H2B with Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade ab52484 at 1/100 dilution (1µg) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labeling MAP2 with Alexa Fluor® 568 Anti-MAP2 antibody [EPR19691] ab303465 at 1/250 dilution (2 μg/ml) (Red). Confocal image showing positive staining in mouse primary neuron. Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection. Anti-MAP2 antibody [HM-2] ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4 μg/ml dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (Green). The Nuclear counterstain was DAPI (Blue).
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neuron cells labeling MAP2 with Alexa Fluor® 568 Anti-MAP2 antibody [EPR19691] ab303465 at 1/250 dilution (2 ug/ml) (Green). Confocal image showing positive staining in rat primary neuron. Confocal scanning Z step was set as 0.3 μm, followed by image processing with maximum Z projection. Anti-MAP2 antibody [HM-2] ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4 μg/ml dilution, followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (Green). The Nuclear counterstain was DAPI (Blue).
MCF7/ HCT 116 cells were fixed in 4% PFA and permeabilized with 0.1% Triton X-100. Primary antibody, Anti-MUC1 antibody [HMFG1 (aka 1.10.F3)] ab70475 at 1:100 was incubated overnight at 4° C, followed by AlexaFluor® 488-conjugated Goat anti-Mouse secondary antibody (ab150113) at 1/1000 dilution at RT for 45 min. Anti-beta Tubulin antibody [EPR16774] ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with Anti-Integrin beta 3 antibody [Y2/51] ab9509 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.
Confocal image showing membranous and cytoplasmic staining in MCF7 cells.
Negative control: HCT 116 (PMID: 14998492)
Flow Cytometry analysis of human monocyte-derived dendritic cells treated with 100ng/mL Lipopolysaccharide (LPS) for 24 hours (Right) / Untreated control (Left).
Anti-CD83 antibody [HB15e] ab123494 used at a 1/1000 dilution.
Secondary is Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) used at a 1/2000 dilution.
The cells were simultaneously stained with CD209. Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
Events were gated on viable single cells.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling CTSD (cathepsin D) with Anti-Cathepsin D [EPR3057Y] - Mouse IgG2a (Chimeric) ab302649 at 1/1000 dilution (0.1ug) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling CTSD (cathepsin D) with Anti-Cathepsin D [EPR3057Y] - Mouse IgG2a (Chimeric) ab302649 at 1/1000 dilution (0.1ug) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.
Flow cytometric analysis of rat bone marrow cells labelling SIRPA with Anti-SIRP alpha antibody [OX-41] ab307563 at 1/1000 dilution (0.1ug) (Right panel) compared with a Mouse monoclonal IgG isotype control (Left panel).
Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.
Gated on viable cells.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PC-3 (human prostate adenocarcinoma epithelial cell) (Left panel) compared to HDLM-2 (human Hodgkin lymphoma cell) (Right panel) cells labelling Semaphorin 4D/CD100 with Anti-Semaphorin 4D/CD100 antibody [30/CD100] ab307685 at 1/1000 dilution (0.1ug) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.
Negative control: PC-3 (PMID: 23775445).
Flow cytometric analysis of HeLa (Human cervix adenocarcinoma epithelial cell) (Left panel) / MCF7 (Human breast adenocarcinoma epithelial cell) (Right panel) cells labeling EpCAM with Anti-EpCAM antibody [AUA1] ab308057 at 1/1000 dilution (0.1ug) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.
Negative control: HeLa. (PMID: 31806375).
Gated on viable cells.
Flow cytometric analysis of 4% paraformaldehyde-fixed 90% methanol-permeabilized RAB8A KO HeLa (ED010027, Left) / Parental HeLa (EDWT01, Right) cells labeling RAB8A with Anti-RAB8A antibody [MJF-R22-79-3] - Mouse IgG2a (Chimeric) ab307607 at 1/800 dilution (0.1 ug) (Red) compared with a mouse monoclonal IgG (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.
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