Suitable for IHC-Fr, ELISA, IHC-P, Flow Cyt, ICC/IF. Ideal for fluorescent cell and tissue imaging. Cited in 3105 publications.
Goat
Rabbit
IgG
Heavy & Light chains
Alexa Fluor® 488
Ex: 495nm, Em: 519nm
IHC-Fr, ELISA, IHC-P, Flow Cyt, ICC/IF
Polyclonal
Application | Reactivity | Dilution info | Notes |
---|---|---|---|
Application IHC-Fr | Reactivity Reacts | Dilution info - | Notes - |
Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
Application IHC-P | Reactivity Reacts | Dilution info - | Notes - |
Application Flow Cyt | Reactivity Reacts | Dilution info 1/2000.00000 - 1/4000.00000 | Notes - |
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, 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
Suitable for IHC-Fr, ELISA, IHC-P, Flow Cyt, ICC/IF. Ideal for fluorescent cell and tissue imaging. Cited in 3105 publications.
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® 488)
Goat
Rabbit
IgG
Heavy & Light chains
Alexa Fluor® 488
Ex: 495nm, Em: 519nm
IHC-Fr, ELISA, IHC-P, Flow Cyt, ICC/IF
Polyclonal
No
This antibody is specific to Rabbit IgG.
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
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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.
Immunoglobulin G (IgG) serves as an essential component of the immune system functioning mainly as an antibody. It is found in blood and extracellular fluid making it one of the most prevalent antibodies in the human body. IgG weighs approximately 150 kDa and it is expressed by B cells as part of the humoral response to pathogens. Alternate names include gamma globulin and Ig gamma. IgG antibodies form a critical aspect of adaptive immunity by targeting specific antigens for neutralization or destruction.
IgG participates in several immune processes by binding to antigens and forming immune complexes. It facilitates pathogen opsonization initiating phagocytosis by immune cells like macrophages and neutrophils. IgG also activates the classical complement pathway which enhances pathogen clearance. IgG exists in a monomeric form and can cross the placenta providing passive immunity to the fetus. In laboratory settings its high specificity makes it valuable in techniques like IgG ELISA for detecting antigen presence.
IgG plays an integral role in the complement system and the adaptive immune response. It partners with proteins such as C1q to initiate the classical complement pathway amplifying the body's ability to detect and remove pathogens. IgG also interacts with Fc receptors on immune cells modulating immune cell activity and facilitating antigen presentation to T cells. These interactions enhance the immune system's efficiency in recognizing and responding to diverse pathogens.
IgG has connections to autoimmune diseases and chronic infections. Elevated levels of IgG can indicate autoimmune conditions like lupus or rheumatoid arthritis where the immune system mistakenly attacks healthy tissue. During chronic infections such as HIV variations in IgG responses can affect disease progression and treatment efficacy. In these contexts IgG interacts with proteins involved in the immune dysregulation characteristic of such diseases highlighting its significant role in both protection and pathology.
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Overlay histogram showing Jurkat cells stained with Anti-CD3 epsilon antibody [SP7] ab16669 (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-CD3 epsilon antibody [SP7] ab16669, 1/1000 dilution) for 30 min at 22°C. The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488) (ab150077) was used at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (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.
Flow cytometric analysis of HEK-293T (Human embryonic kidney epithelial cell, Left panel) / Caco-2 (Human colorectal adenocarcinoma epithelial cell, Right panel) using Anti-CD133 antibody [RM1002] - Stem Cell Marker ab278053 at 1/500 dilution (0.1µg) (red). The isotype control used was the Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730), black line and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Secondary antibody used was the Goat anti-rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution.
Negative control: HEK-293T. (PMID 20167130)
Gated on viable cells.
ICC/IF image of beta Tubulin staining in 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 primary antibody (Anti-beta Tubulin antibody - Loading Control ab6046, 5μg/ml) overnight at +4°C. The secondary antibody (green) was ab150077 Alexa Fluor® 488 goat anti-rabbit 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 - Wholemount of Caenorhabditis elegans larvae labelling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody ab5095. The sample was incubated with primary antibody (1/500 in PBS + 3% BSA + 0.1% Triton X-100) for 12 hours at 4°C. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/1000), was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence analysis of C6(Rat glial tumor glial cell) cells labeling alpha smooth muscle Actin with purified Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] ab32575 at 1/100 dilution (0.71 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Immunocytochemistry analysis of SK-OV-3 (human ovarian cancer epithelial cell)cells labelling Zic2 with Anti-Zic2 antibody [EPR7790] ab150404 at 2.5 μg/ml. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 was used as the secondary antibody (green). Cells were counterstained with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) at 1/200 dilution (red). Nuclear DNA was labelled with DAPI (blue).
Immunohistochemistry (Frozen sections) analysis of rat stomach tissue (4% PFA, 0.2% Tritron x-100) labelling Integrin alpha 6 with Anti-Integrin alpha 6 antibody [EPR18124] - BSA and Azide free ab240252 at 1/100 dilution. ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/1000 dilution. Negative control using PBS instead of primary antibody (left). Counterstained with DAPI.
Positive staining was seen on rat stomach.
Overlay histogram showing HeLa cells stained with Anti-Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade ab32356 (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-Histone H3 (di methyl K4) antibody [Y47] - ChIP Grade ab32356, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/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.
Overlay histogram showing MCF7 cells stained with unpurified Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade ab32063 (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-Estrogen Receptor alpha antibody [E115] - ChIP Grade ab32063, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (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. This antibody gave a positive signal in MCF7 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3(Mouse embryonic fibroblast) cells labeling alpha smooth muscle Actin with purified Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] ab32575 at 1/500 dilution (5.2 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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. 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 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).
Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) treated with 25 ug/mL anisomycin for 30 min, then Lambda Protein Phosphatase 31 for 2 hours cells labeling Hsp27 with purified Anti-Hsp27 (phospho S78) antibody [Y175] ab32501 at 1/50 dilution (11.3 μg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 μg/mL) dilution. Goat anti-rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Immunocytochemistry analysis of Jurkat (human T cell leukemia T lymphocyte) labeling p95/NBS1 with purified Anti-p95/NBS1 (phospho S343) antibody [EP178] ab109453 at 1/100 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 μg/ml) was used as the secondary antibody. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.27 μg/ml) was used as counterstain. Nuclei were stained blue with DAPI. Negative control: PBS instead of the primary antibody.
Confocal image showing increased nuclear staining in Jurkat cells treated with Etoposide (25uM) for 2 h.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling KDM3B / JMJD1B with Anti-KDM3B / JMJD1B antibody [EPR24219-30] ab271046 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HeLa cell line. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubμue Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
Immunocytochemistry/Immunofluorescence analysis of Raji (human Burkitt's lymphoma) cells labelling HLA A with purified Anti-HLA A antibody [EP1395Y] ab52922 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling LAMP1 with Anti-LAMP1 antibody [EPR24395-31] ab278043 at 1/100 (5.45 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 (green). Confocal image showing cytoplasmic staining in HeLa. Anti-LAMP1 antibody [H4A3] ab25630 Anti-LAMP1 mouse monoclonal antibody was used to counterstain at 1/500 (red). The Nuclear counterstain was DAPI (blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution
Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling ATG9A with Purified Anti-ATG9A antibody [EPR2450(2)] ab108338 at 1/100 dilution. Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) was used as the secondary antibody at 1/1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 cells labeling KDM3B / JMJD1B with Anti-KDM3B / JMJD1B antibody [EPR24219-30] ab271046 at 1/500 dilution, followed by 150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in NIH/3T3 cell line. 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 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
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. 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 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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 (ab150077).
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) and (Anti-Lamin B1 antibody - Nuclear Envelope Marker ab16048, 1μg/ml) overnight at +4°C. The secondary antibodies were Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) ab150115 Alexa Fluor® 647 (red) goat anti-mouse IgG (H+L) used at 2μg/ml for 1h and ab150077 Alexa Fluor® 488 (green) goat anti-rabbit IgG (H+L) used at 2μg/ml for 1h. DAPI was used to stain the cell nuclei.
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 LAMP1 KO HAP1 cells labelling LAMP1 with Anti-LAMP1 antibody [EPR24395-31] ab278043 at 1/100 (5.45 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 ug/ml dilution (green). Confocal image showing cytoplasmic staining in parental HAP1 cell line and no staining in LAMP1 KO HAP1 cell line. 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 at 1/200 (red).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
Intracellular Flow Cytometry analysis of 293T (human embryonic kidney epithelial cell) transfected with myc-tagged PADI4 construct, then treated with 10mM CaCl2 and 10uM Ionomycin for 4h labeling Histone H3 (citrulline R2 + R8 + R17) with Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] ab281584 at 1/500 dilution. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as a secondary antibody.
Isotype control : Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730)/left.
Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling CD2 with purified Anti-CD2 antibody [EPR6451] ab131276 at 1:20 dilution (10 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as a isotype control. Cell without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.
Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling CSNK2A1 with purified Anti-CSNK2A1 antibody [EP1963Y] ab76040 at 1:20 dilution (10 μg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as a isotype control. Cell without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.
Immunocytochemistry/ Immunofluorescence analysis of A549 (Human lung carcinoma epithelial cell) cells labeling OGT / O-Linked N-Acetylglucosamine Transferase using Anti-OGT / O-Linked N-Acetylglucosamine Transferase antibody [EPR12713] ab177941. The cells were fixed with 4% paraformaldehyde then permeabilized with 0.1% Triton X-100. The cells were then incubated with Anti-OGT / O-Linked N-Acetylglucosamine Transferase antibody [EPR12713] ab177941 at 1:100 dilution followed by a further incubation with a Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI. Cells were counterstained using Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1:200 dilution (shown in red). Secondary antibody only control: PBS instead of the primary antibody.
Immunocytochemistry/ Immunofluorescence analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling NDRG1 using Anti-NDRG1 antibody [EPR5593] ab124689. The cells were fixed with 100% Methanol then permeabilized with 0.1% Triton X-100. The cells were then incubated with Anti-NDRG1 antibody [EPR5593] ab124689 at 1:50 dilution followed by a further incubation with a Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI. Cells were counterstained using Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1:200 dilution (shown in red). Secondary antibody only control: PBS instead of the primary antibody.
Immunocytochemistry/ Immunofluorescence analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labeling S100 alpha 2 using Anti-S100 alpha 2/S100A2 antibody [EPR5392] ab109494. The cells were fixed with 100% Methanol then permeabilized with 0.1% Triton X-100. The cells were then incubated with Anti-S100 alpha 2/S100A2 antibody [EPR5392] ab109494 at 1:100 dilution followed by a further incubation with a Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI. Cells were counterstained using Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1:200 dilution (shown in red). Secondary antibody only control: PBS instead of the primary antibody.
Immunocytochemistry/ Immunofluorescence analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labeling FRA1 using Anti-FRA1 antibody [EP4711] ab124722. The cells were fixed with 4% paraformaldehyde then permeabilized with 0.1% Triton X-100. The cells were then incubated with Anti-FRA1 antibody [EP4711] ab124722 at 1:50 dilution followed by a further incubation with a Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI. Cells were counterstained using Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1:200 dilution (shown in red). Secondary antibody only control: PBS instead of the primary antibody.
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling PKC beta 2 using Anti-PKC beta 2 antibody [Y125] ab32026. The cells were fixed with 4% paraformaldehyde then permeabilized with 0.1% Triton X-100. The cells were then incubated with Anti-PKC beta 2 antibody [Y125] ab32026 at 1:50 dilution followed by a further incubation with a Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI. Cells were counterstained using Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1:200 dilution (shown in red). Secondary antibody only control: PBS instead of the primary antibody.
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling IMP3 using Anti-IMP3 antibody [EPR12021] ab177477. The cells were fixed with 4% paraformaldehyde then permeabilized with 0.1% Triton X-100. The cells were then incubated with Anti-IMP3 antibody [EPR12021] ab177477 at 1:50 dilution followed by a further incubation with a Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI. Cells were counterstained using Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1:200 dilution (shown in red). Secondary antibody only control: PBS instead of the primary antibody.
Immunocytochemistry/ Immunofluorescence analysis of PBMC (Human primary peripheral blood mononuclear cell) cells labeling IL-2 using Anti-IL-2 antibody [EPR2780] ab92381. The cells were fixed with 4% paraformaldehyde then permeabilized with 0.1% Triton X-100. The cells were then incubated with Anti-IL-2 antibody [EPR2780] ab92381 at 1:50 dilution followed by a further incubation with a Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI. Cells were counterstained using Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1:200 dilution (shown in red). Secondary antibody only control: PBS instead of the primary antibody.
Anti-Collagen XVII antibody [EPR14758] ab186415 staining Collagen XVII in A431 (Human epidermoid carcinoma epithelial cell) cells. The cells were fixed with 4% paraformaldehyde then permeabilized with 0.1% tritonX-100. The cells were then incubated with Anti-Collagen XVII antibody [EPR14758] ab186415 at 10μg/ml concentration followed by a further incubation with a Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used at 1/200 dilution as counterstain (shown in red). Secondary antibody only control: PBS instead of the primary antibody.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (Human embryonic kidney epithelial cell) transfected with HA tagged IL-21 construct cells labelling Il21 with Anti-IL-21 antibody [EPR22618-28] ab227837 at 1/500 dilution (0.1ug)/ Right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized CTNNB1 KO HAP1 (CTNNB1 knockout human chronic myelogenous leukemia near-haploid cell line) cells labelling beta Catenin non-phospho S37/T44 with Anti-beta Catenin non-phospho S37/T41 antibody [EPR23969-131] ab246504 at 1/100 (5.4 μg/ml) dilution, followed by ab150077 antibody at 1/1000 (2 μg/ml) dilution (Green). Confocal image showing membranous staining in parental HAP1 cell line, and staining in the CTNNB1 KO HAP1 cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 was used to counterstain tubulin at 1/200 (2.5 μg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 at 1/1000 (2 μg/ml) dilution.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Th17 polarized Mouse splenocytes were treated with 500ng/ml PMA and 500 ng/ml ionomycin for 1 hours, then add 1ug/ml Brefeldin A for another 4 hours cells labelling Il21 with Anti-IL-21 antibody [EPR22618-28] ab227837 at 1/500 dilution (0.1ug)/ Right (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. IL21 is mutual exclusively expressed with IFN gamma (Th1 marker).Cells were provided by an anonymous collaborator.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-101 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling beta Catenin non-phospho S37/T45 with Anti-beta Catenin non-phospho S37/T41 antibody [EPR23969-131] ab246504 at 1/100 (5.4 μg/ml) dilution, followed by ab150077 antibody at 1/1000 (2 μg/ml) dilution (Green). Confocal image showing membranous and cytoplasmic staining in NIH/3T3 cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 was used to counterstain tubulin at 1/200 (2.5 μg/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 555) ab150078 at 1/1000 (2 μg/ml) dilution.
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