Goat anti-rabbit IgG H&L (Alexa Fluor® 594) is a 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 ICC/IF, IHC, flow cytometry and ELISA.
- 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 650 publications
Application | Reactivity | Dilution info | Notes |
---|---|---|---|
Application IHC-Fr | Reactivity Reacts | Dilution info - | Notes - |
Application ICC/IF | Reactivity Reacts | Dilution info 1/200 - 1/1000 | Notes - |
Application Flow Cyt | Reactivity Reacts | Dilution info 1/2000 - 1/4000 | Notes - |
Application IHC-P | Reactivity Reacts | Dilution info - | Notes Use at an assay dependent dilution |
Application ELISA | Reactivity Reacts | Dilution info - | Notes Use at an assay dependent dilution |
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Ig gamma 1 chain C region, Ig gamma 3 chain C region, Ig gamma 4 chain C region, Ig gamma chain C region, Ig gamma-2 chain C region, Ig kappa-b4 chain C region, Ig kappa-b5 chain C region, Ig kappa-b9 chain C region, Ig lambda chain C region, Immunoglobin heavy constant gamma 1, Immunoglobulin G, K-BAS
Goat anti-rabbit IgG H&L (Alexa Fluor® 594) is a 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 ICC/IF, IHC, flow cytometry and ELISA.
- 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 650 publications
Preservative: 0.02% Sodium azide
Constituents: PBS, 23% Glycerol (glycerin, glycerine), 1% BSA
The antibody was isolated by affinity chromatography using antigen coupled to agarose beads.
Fluorochrome chart- a complete quick and easy guide to help you select the most appropriate fluorochromes for your next experiment.
When it comes to advancing your immunohistochemistry (IHC) research, Abcam offers a comprehensive suite of IHC kits and secondary antibodies, designed to deliver precise and reliable results.
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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Overlay histogram showing Jurkat cells stained with Anti-CD45 antibody [EP322Y] ab40763 (red line). The cells were fixed with 4% paraformaldehyde (10 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-CD45 antibody [EP322Y] ab40763, 1/1000 dilution) for 30 min at 22°C. The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 594) (ab150080) was used at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730, 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 561nm laser and 610/20 bandpass filter.
ICC/IF image of Anti-beta Tubulin antibody - Loading Control ab6046 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then incubated in 1%BSA / 10% normal donkey 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 primary antibody (Anti-beta Tubulin antibody - Loading Control ab6046, 5μg/ml) overnight at +4°C. The secondary antibody (orange) was ab150080 Alexa Fluor® 4594 goat anti-rabbir 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.
IHC-P image of alpha smooth muscle actin (Anti-alpha smooth muscle Actin antibody ab5694) staining E16.5 mouse embryo gut. Paraformaldehyde fixed and paraffin embedded E16.5 mouse embryo gut sections were dewaxed and rehydrated before antigen retrieval (4 mins in a pressure cooker in 10mM Tris/0.4mM EDTA buffer pH 9.5). They were then incubated in 50mM NH4Cl for 30 minutes and washed/blocked in 3x 10 minute washes of PBS containing 1% BSA + 0.2% gelatine and 0.05% saponin. Sections were incubated overnight with a primary antibody against alpha smooth muscle actin (Anti-alpha smooth muscle Actin antibody ab5694), diluted 1/250 in PBS containing 0.1% BSA and 0.3% triton. After 3 x 10 minute washes in of PBS containing 0.1% BSA, 0.2% gelatine and 0.05% saponin, the sections were incubated for 1 hr in the secondary antibody (ab150080, diluted 1/400, shown in red) and then the 3 washes repeated. Sections were mounted in Vectashield with DAPI (blue).
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 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Anti-beta Tubulin antibody [EPR16774] - Loading Control ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution followed by 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 ab150080 at a 1/1000 dilution.
Negative control 2: Anti-beta Tubulin antibody [EPR16774] - Loading Control ab179513 at a 1/200 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 at a 1/1000 dilution.
Cross-reactivity of the polyclonal secondary antibody Goat Anti-Rabbit IgG H&L ab182016 was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards at 1 μg/ml (50 μl/well) and incubated overnight at 4°C, followed by a 5% BSA blocking step for 2h at RT. Goat Anti-Rabbit IgG H&L ab182016 was then added starting at 1 μg/ml and gradually diluted 1/4 (50 μl/well), followed by incubation for 2h. For the detection Donkey anti-Goat IgG H&L (HRP) (Donkey Anti-Goat IgG H&L (HRP) ab6885) was used at 1/10,000 dilution (50 μl/well), followed by incubation for 1h at RT.
For the batch tested, Goat Anti-Rabbit IgG H&L ab182016 showed a cross-reactivity of 5-7% towards Human IgG and below 2% towards Mouse IgG, Rat IgG and Chicken IgY.
This data was developed using the unconjugated antibody (Goat Anti-Rabbit IgG H&L ab182016).
Cross-reactivity of Goat anti-Rabbit IgG H&L (Goat Anti-Rabbit IgG H&L ab182016) and Goat anti-Rabbit IgG H&L obtained from two different vendors was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards (Rabbit, Human, Mouse and Rat) at 1 μg/ml (50 μl/well) and incubated overnight at 4°C, followed by a 5% BSA blocking step for 2h at RT. Secondary antibodies were then added starting at 1 μg/ml and gradually diluted 1/4 (50 μl/well), followed by incubation for 2h. For the detection Donkey anti-Goat IgG H&L (HRP) (Donkey Anti-Goat IgG H&L (HRP) ab6885) was used at 1/10,000 dilution (50 μl/well), followed by incubation for 1h at RT. This data is from a representative dilution.
This data was developed using the unconjugated antibody (Goat Anti-Rabbit IgG H&L ab182016).
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (Human embryonic kidney epithelial cell) transfected with His and Myc tagged cas9 construct cells labelling 6X His tag with Anti-6X His tag® antibody [AD1.1.10] ab15149 at 1/100 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution(Green).
Confocal image showing cytoplasmic staining in 293T cells transfected with His and Myc tagged mouse cas9 construct. The counterstain was observed in red. Nuclear DNA was labeled with DAPI (shown in blue).
Anti-c-Myc antibody [Y69] - ChIP Grade ab32072 Anti-c-Myc rabbit monoclonal antibody was used to counterstain Myc at 1/500 followed by a secondary antibody ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at a 1/1000 dilution (Red).
Immunofluorescence staining of Doublecortin in a section of frozen PFA perfusion fixed 6 week old rat dentate gyrus.
After dissection, the tissue was further fixed in PFA overnight, washed in PBS and then incubated overnight in 30% sucrose. It was then embedded in OCT and cryosectioned. No antigen retrieval step was performed prior to staining. Performed on a Leica BONDTM. The section was incubated at room temperature for 1 hour with Anti-Doublecortin antibody ab18723 at 1ug/mL and then incubated with ab150080 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preabsorbed, (Shown in magenta) 1/1000) for 1 hour at room temperature. Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized mouse primary neural/glia cells labelling Myelin Basic Protein with Anti-Myelin Basic Protein antibody [12] ab7349 at a 1/200 dilution, followed by Goat anti-Rat (Alexa Fluor® 488) (Goat Anti-Rat IgG H&L (Alexa Fluor® 488) ab150157) secondary antibody at a 1/1000 dilution. Confocal image showing positive staining in mouse primary oligodendroglia cells (shown in green). Nuclear DNA was labeled with DAPI (shown in blue).
Counterstained with Anti-MAP2 antibody (Anti-MAP2 antibody [EPR19691] - Neuronal Marker ab183830) at a 1/1000 dilution, followed by Goat Anti-Rabbit secondary (Alexa Fluor® 594) (ab150080) at a 1/1000 dilution (shown in magenta).
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).
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neuron cells labelling Glutamate Receptor 1 (AMPA subtype) with Anti-Glutamate Receptor 1 (AMPA subtype) antibody [N355/1] - N-terminal ab174785 at 1/200 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 1/1000 dilution at RT for 45 min. Anti-MAP2 antibody [EPR19691] (Anti-MAP2 antibody [EPR19691] - Neuronal Marker ab183830) was used as a counterstain at 1/1000 dilution and was co-incubated with Anti-Glutamate Receptor 1 (AMPA subtype) antibody [N355/1] - N-terminal ab174785 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Rat primary neuron cells labelling Glutamate Receptor 1 (AMPA subtype) with Anti-Glutamate Receptor 1 (AMPA subtype) antibody [N355/1] - N-terminal ab174785 at 1/200 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 1/1000 dilution at RT for 45 min. Anti-MAP2 antibody [EPR19691] (Anti-MAP2 antibody [EPR19691] - Neuronal Marker ab183830) was used as a counterstain at 1/1000 dilution and was co-incubated with Anti-Glutamate Receptor 1 (AMPA subtype) antibody [N355/1] - N-terminal ab174785 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A375 (human malignant melanoma epithelial cell) cells labelling SOX10 with Anti-SOX10 antibody [SOX10/991] ab218522 at 1/50 dilution followed by secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (green).
Anti-beta Tubulin antibody [EPR16774] - Loading Control ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used as a counterstain at a 1/200 dilution followed by a secondary antibody ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at a 1/1000 dilution (red).
Confocal image showing nuclear staining in A375 cells.
Negative control: MCF7 (PMID: 23799842).
Nuclear DNA was labelled with DAPI (shown in blue).
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized A431 cells labelling EGFR with Anti-EGFR antibody [ICR10] ab231 at 1/50 dilution followed by secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (green).
Anti-beta Tubulin antibody [EPR16774] - Loading Control ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used as a counterstain at a 1/200 dilution followed by a secondary antibody ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) used at a 1/1000 dilution (red).
Confocal image showing strong membranous and weak cytoplasmic staining in A431 cells.
Low expression control: HEK-293 (PMID: 26368334).
Nuclear DNA was labelled with DAPI (shown in blue).
Confocal image showing membranous staining in RAW 264.7 cells.
RAW 264.7, WEHI-231 and THP-1 cells were fixed in 4% PFA and permeabilized with 0.1% Triton X-100. Primary antibody, Anti-CD11b antibody [M1/70] ab8878 at 1:50 was incubated overnight at 4° C, followed by AlexaFluor® 488- conjugated Goat anti-Rat secondary antibody (Goat Anti-Rat IgG H&L (Alexa Fluor® 488) ab150157) at 1/1000 dilution at RT for 45 min. Anti-beta Tubulin antibody [EPR16774] - Loading Control ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with Anti-CD11b antibody [M1/70] ab8878 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.
Negative control: WEHI-231 (PMID: 2457584)
RAW 264.7 cells were fixed in 4% PFA and permeabilized with 0.1% Triton X-100. Primary antibody, Anti-CD68 antibody [FA-11] ab53444 at 1:50 was incubated overnight at 4° C, followed by AlexaFluor® 488-conjugated Goat anti-Rat secondary antibody (Goat Anti-Rat IgG H&L (Alexa Fluor® 488) ab150157) at 1/1000 dilution at RT for 45 min. Anti-beta Tubulin antibody [EPR16774] - Loading Control ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with Anti-CD68 antibody [FA-11] ab53444 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI. Confocal image showing cytoplasmic staining in RAW 264.7 cell line.
NIH/3T3 cells were fixed in 100% methanol and permeabilized with 0.1% Triton X-100. Primary antibody, Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade ab52484 at 1:2000 was incubated overnight at 4° C, followed by AlexaFluor® 488-conjugated Goat anti-mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 1/1000 dilution at RT for 45 min. Anti-beta Tubulin antibody [EPR16774] - Loading Control ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade ab52484 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 cells labelling CEACAM1 + CEACAM3 + CEACAM6 with Anti-CEACAM1 + CEACAM3 + CEACAM6 antibody [YTH71.3] ab104450 at 1/250 dilution followed by secondary antibody Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (green).
Anti-beta Tubulin antibody [EPR16774] - Loading Control ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used as a counterstain at a 1/200 dilution followed by a secondary antibody ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) used at a 1/1000 dilution (red).
Confocal image showing membranous staining in HepG2 cell line.
Negative control: HeLa (PMID: 11580753).
Nuclear DNA was labelled with DAPI (shown in blue).
Anti-HIF-1 alpha antibody [H1alpha67] ab1 staining HIF-1 alpha in HeLa DFO 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-HIF-1 alpha antibody [H1alpha67] ab1 at 10µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Anti-HIF-1 alpha antibody [ESEE122] ab8366 staining HIF-1 alpha in Hela cells. Untreated and BrdU treated (10µM for 24 hours) cells. The cells were fixed with 4% paraformaldehyde (10 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-HIF-1 alpha antibody [ESEE122] ab8366 at 10µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) 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.
HeLa cells were fixed in 100% methanol and permeabilized with 0.1% Triton X-100. Primary antibody, Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade ab52484 at 1:2000 was incubated overnight at 4° C, followed by AlexaFluor® 488-conjugated Goat anti-mouse secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 1/1000 dilution at RT for 45 min. Anti-beta Tubulin antibody [EPR16774] - Loading Control ab179513 Anti-beta Tubulin, used as a counterstain at 1/200 dilution, was co-incubated with Anti-Histone H2B antibody [mAbcam 52484] - ChIP Grade ab52484 overnight at 4° C, followed by Alexa Fluor® 594 Goat Anti-Rabbit secondary (ab150080) at 1/1000 dilution at RT for 45 min. Nucleus were visualized using DAPI.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Human papillomavirus L2 protein with Anti-Human papillomavirus L2 antibody [K18_HPV16L2(20-38)] ab322522 at 1/100 (10.45 ug/ml) dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).
Confocal image showing nuclear staining in 293T cells transfected with a Human papillomavirus type 16 L2 protein expression vector containing a myc-His-tag®(shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-Myc tag antibody ab9106 Anti-Myc Rabbit polyclonal antibody was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/500 (4μg/ml) dilution (Magenta).
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling Human papillomavirus L2 protein with Anti-Human Papilloma Virus L2 antibody [K4_HPV16L2(20-38)] ab321817 at 1/2000 (0.51 ug/ml) dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).
Confocal image showing nuclear and cytoplasmic staining in 293T cells transfected with a Human papillomavirus type 16 L2 protein expression vector containing a myc-His-tag®(shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-Myc tag antibody ab9106 Anti-Myc Rabbit polyclonal antibody was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/500 (4 ug/ml) dilution (Magenta).
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Raw 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling groEL with Anti-groEL antibody [EPR28718-8] ab318970 at 1/2000 (0.251 ug/ml) dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) antibody at 1/500 4ug/ml dilution (Green).
Confocal image showing positive staining in Raw 264.7 cells (shown in magenta) infected by Escherichia coli BL21(DE3), which was transformed with an eGFP expression vector. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
was used to counterstain tubulin at 1/200 2.5 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/500 4ug/ml dilution.
Anti-Vimentin antibody [LN-6] ab230171 staining Vimentin in HeLa-VIM cells. The cells were fixed with 4% paraformaldehyde (10 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-Vimentin antibody [LN-6] ab230171 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046. Cells were then incubated with Goat Anti-Mouse IgM mu chain (Alexa Fluor® 488) ab150121 at 1/1000 dilution (shown in green) and ab150080 at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 100% methanol (5 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Anti-A2B5 antibody [105] ab53521 staining A2B5 in primary mouse neurons/glia, DIV14 (prepared from E18 mouse hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. C57EHP) cells. The cells were fixed with 4% paraformaldehyde (10 min), 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 1h. The cells were then incubated overnight at 4°C with Anti-A2B5 antibody [105] ab53521 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046. Cells were then incubated with Goat Anti-Mouse IgM mu chain (Alexa Fluor® 488) ab150121 at 1/1000 dilution (shown in green) and ab150080 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.
Anti-A2B5 antibody [105] ab53521 staining A2B5 in primary rat neurons/glia, DIV14 (prepared from E18 rat hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. SDHEP) cells. The cells were fixed with 4% paraformaldehyde (10 min), 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 1h. The cells were then incubated overnight at 4°C with Anti-A2B5 antibody [105] ab53521 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046. Cells were then incubated with Goat Anti-Mouse IgM mu chain (Alexa Fluor® 488) ab150121 at 1/1000 dilution (shown in green) and ab150080 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.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney) transfected with GFP-tagged human HA Tag expression vector cells labelling HA tag with Anti-HA tag antibody [RM1058] ab314237 at 1/500 (1.042 ug/ml) dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) antibody at 1/1000 (2 ug/mL) dilution (Green).
Confocal image showing cytoplasmic staining in 293T cells transfected with a human HA Tag expression vector containing a GFP tag.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).
The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) at 1/1000 (2 ug/mL) dilution.
Anti-Cytokeratin 15 antibody [LHK15] ab80522 staining Cytokeratin 15 in A431 cells. The cells were fixed with 4% paraformaldehyde (10 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-Cytokeratin 15 antibody [LHK15] ab80522 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 100% methanol (5 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Anti-Histone H2B antibody [mAbcam 64165] - ChIP Grade ab64165 staining Histone H2B in 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-Histone H2B antibody [mAbcam 64165] - ChIP Grade ab64165 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) 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.
Anti-HOXB13 antibody [mAbcam53931] ab53931 staining HOXB13 in HepG2 cells. The cells were fixed with 4% paraformaldehyde (10 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-HOXB13 antibody [mAbcam53931] ab53931 at 5µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) 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.
Anti-Histone H4 antibody [mAbcam 31830] - ChIP Grade ab31830 staining Histone H4 in 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-Histone H4 antibody [mAbcam 31830] - ChIP Grade ab31830 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Anti-CD32B + CD32A antibody [AT10] ab23336 staining CD32B + CD32A in K-562 cells. The cells were fixed with 4% paraformaldehyde (10 min), 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 1h. The cells were then incubated overnight at 4°C with Anti-CD32B + CD32A antibody [AT10] ab23336 at 5µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour magenta). 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..
Anti-Integrin alpha V+beta 5 antibody [P1F6] ab177004 staining Integrin alpha V+beta 5 in A549 cells. The cells were fixed with 4% paraformaldehyde (10 min), 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 1h. The cells were then incubated overnight at 4°C with Anti-Integrin alpha V+beta 5 antibody [P1F6] ab177004 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) 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.
Anti-Histone H3 (phospho S10) antibody [mAbcam 14955] - BSA and Azide free ab238673 staining Histone H3 (phospho S10) in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 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-Histone H3 (phospho S10) antibody [mAbcam 14955] - BSA and Azide free ab238673 at 5µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 100% methanol (5 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Anti-Cytochrome C antibody [7H8.2C12] ab13575 staining Cytochrome C in HepG2 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-Cytochrome C antibody [7H8.2C12] ab13575 at 1µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Anti-MiTF antibody [C5] ab12039 staining MiTF in Malme-3M cells. The cells were 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-MiTF antibody [C5] ab12039 at 5µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) 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.
Anti-WIPI2 antibody [2A2] ab105459 staining WIPI2 in HeLa Chloroquine treated 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-WIPI2 antibody [2A2] ab105459 at 5µg/ml and Anti-beta Tubulin antibody - Loading Control ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L Alexa Fluor® 488) preadsorbed at 1/1000 dilution shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L Alexa Fluor® 594) 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.
Anti-DDX4 / MVH antibody [mAbcam27591] ab27591 staining DDX4/MVH in mES cells. The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% 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 with Anti-DDX4 / MVH antibody [mAbcam27591] ab27591 at 10μg/ml concentration and Anti-beta Tubulin antibody - Loading Control ab6046 (Rabbit polyclonal to beta Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to mouse IgG (Alexa Fluor® 488) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 2 μg/ml (shown in green) and a goat secondary antibody to rabbit IgG (Alexa Fluor® 594) (ab150080) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).
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