Name: Anti-GAPDH antibody [EPR6256] - Loading Control
SKU: ab128915
Rating: 5 (11 reviews)

Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915) is a rabbit monoclonal antibody detecting GAPDH in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF. Suitable for African green monkey, Human.

- Biophysical QC for unrivalled batch-batch consistency

- Over 270 publications

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ab221270
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PerCP Anti-GAPDH antibody [EPR6256]
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Alexa Fluor® 647 Anti-GAPDH antibody [EPR6256]
ab195904
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Biotin Anti-GAPDH antibody [EPR6256] - Loading Control
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HRP Anti-GAPDH antibody [EPR6256] - Loading Control
ab186930
Carrier free
Anti-GAPDH antibody [EPR6256] - BSA and Azide free

Images

Western blot - Anti-GAPDH antibody [EPR6256] - Loading Control (AB128915), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested	Tested
African green monkey	Expected	Tested	Expected	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/10 - 1/100	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species African green monkey	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species African green monkey	Dilution info 1/10000 - 1/50000	Notes -
Species Human	Dilution info 1/10000 - 1/50000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/250 - 1/500	Notes 2.0 μg/ml

Expected
Expected

Species	Dilution info	Notes
Species African green monkey	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/20	Notes For unpurified use at 1/100 - 1/1000. Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species	Dilution info	Notes
Species African green monkey	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/2000	Notes For unpurified use at 1/250. The African green monkey recommendation is based on the WB results. We do not guarantee IHC-P for African green monkey. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species African green monkey	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

Select an associated product type

15 products for Alternative Product

Target data

See full target information GAPDH

Function

Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively (PubMed:11724794, PubMed:3170585). Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate (PubMed:11724794, PubMed:3170585). Modulates the organization and assembly of the cytoskeleton (By similarity). Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules (By similarity). Component of the GAIT (gamma interferon-activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes (PubMed:23071094). Upon interferon-gamma treatment assembles into the GAIT complex which binds to stem loop-containing GAIT elements in the 3'-UTR of diverse inflammatory mRNAs (such as ceruplasmin) and suppresses their translation (PubMed:23071094). Also plays a role in innate immunity by promoting TNF-induced NF-kappa-B activation and type I interferon production, via interaction with TRAF2 and TRAF3, respectively (PubMed:23332158, PubMed:27387501). Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis (By similarity). Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC (By similarity).

Alternative names

GAPD, CDABP0047, OK/SW-cl.12, GAPDH, Glyceraldehyde-3-phosphate dehydrogenase, Peptidyl-cysteine S-nitrosylase GAPDH

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR6256
Purification technique: Affinity purification Protein A
Specificity: The African green monkey recommendation is based on the WB results. We do not guarantee IHC-P for African green monkey.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle, Stable for 12 months at -20°C

Notes

What is this antibody validated in?

Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF) in African green monkey, Human samples.

What is the molecular weight of GAPDH?

Anti-GAPDH [EPR6256] - Loading Control (ab128915) is often used as a loading control in Western blot. It specifically detects a band for GAPDH (UniProt: P04406) at a molecular weight of 36kDa.

Trusted by the scientific community

Anti-GAPDH [EPR6256] - Loading Control (ab128915) was first used in a scientific publication in 2012 and has been cited over 270 times in peer-reviewed journals.

Reviewed by scientists

Anti-GAPDH [EPR6256] - Loading Control (ab128915) has over 10 independent reviews from customers.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Other related products

We have a range of other formats of antibody clone [EPR6256] also available for your convenience:
ab128915, HRP - ab185059, Carrier free - ab186930, Biotin - ab195904, Alexa Fluor® 647 - ab215227, PerCP - ab216377, APC - ab221270

Species reactivity
Mouse: We have preliminary internal testing data to indicate this antibody may not react with this species.
Please contact us for more information.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Glyceraldehyde-3-phosphate dehydrogenase commonly known as GAPDH is an enzyme involved in glycolysis. Its molecular weight (MW) is approximately 36 kDa. The protein is expressed ubiquitously in almost all tissues reflecting its essential role in energy production. GAPDH catalyzes the sixth step of glycolysis converting glyceraldehyde-3-phosphate into 13-bisphosphoglycerate. Due to its stable expression researchers often use GAPDH as a loading control in western blot experiments.

Biological function summary

GAPDH serves important metabolic functions beyond its enzymatic role in glycolysis. It functions as part of a multi-enzyme complex within the cytoplasm which facilitates efficient substrate channeling during glycolysis. Additionally GAPDH has non-glycolytic roles including involvement in nuclear processes like RNA export and DNA repair. Its ubiquitous presence across different cellular compartments indicates its multiple functions beyond metabolic pathways.

Pathways

GAPDH integrates into significant cellular functions like the glycolytic pathway and apoptotic pathways. In glycolysis GAPDH collaborates with enzymes like phosphoglycerate kinase forming a cohesive link in the energy conversion chain. Its participation in apoptotic pathways highlights GAPDH's involvement in cellular death processes interacting with proteins like Bcl-2 to influence apoptosis progression. These roles reinforce its presence in central metabolic and regulatory pathways.

Associated diseases and disorders

GAPDH has associations with neurodegenerative diseases and cancer. In neurodegenerative disorders such as Alzheimer's disease GAPDH’s altered enzymatic activity is frequently observed influencing cellular energy homeostasis. Moreover overexpression or aberrant regulation of GAPDH relates to cancer cell proliferation and metastasis implicating proteins like p53 in these pathways. The diverse functions and interactions of GAPDH emphasize its importance in both normal cellular function and disease states.

Product promise

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11 product images

Western blot - Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915)
All lanes: Western blot - Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915) at 1/20000 dilution
All lanes: COS-1 (African green monkey kidney fibroblast-like) whole cell lysates at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa
Western blot - Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915)
All lanes: Western blot - Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915) at 1/20000 dilution
All lanes: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa
This image is courtesy of a customer review submitted by Kirk McManus
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915)
ab128915 (unpurified) staining GAPDH in human HeLa (human epithelial cell line from cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X-100 in PBS. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody. Counter stained with DAPI.
Western blot - Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915)
Secondary antibody - anti-rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab6721)
All lanes: Western blot - Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915) at 1/10000 dilution
Lane 1: HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate at 10 µg
Lane 2: HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate at 10 µg
Lane 3: HepG2 (human liver hepatocellular carcinoma cell line) cell lysate at 10 µg
Lane 4: HUVEC (human umbilical vein endothelial cell line) cell lysate at 10 µg
Lane 5: MCF7 (human breast adenocarcinoma cell line) cell lysate at 10 µg
Lane 6: SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate at 10 µg
Secondary
All lanes: HRP labelled Goat anti-Rabbit IgG at 1/2000 dilution
Predicted band size: 36 kDa
Observed band size: 35 kDa
Immunoprecipitation - Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915)
ab128915 (purified) at 1/20 dilution (0.5ug) immunoprecipitating GAPDH in HeLa whole cell lysates.
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates 10ug
Lane 2 (+): ab128915 & HeLa whole cell lysates
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab128915 in HeLa whole cell lysates
For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915)
Predicted band size: 25 kDa, 36 kDa
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915)
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling GAPDH with purified ab128915 at 1/250 dilution (0.4 μg/ml). Cells were fixed in 100% Methanol. 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, Goat Anti-Rabbit IgG H&L (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.
Flow Cytometry (Intracellular) - Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915)
Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling GAPDH with purified ab128915 at 1/20 dilution (10μg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human bladder carcinoma tissue sections labeling GAPDH with purified ab128915 at 1/2000 dilution (0.06 µg/ml). Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915)
ab128915 (unpurified) staining GAPDH in HeLa (human epithelial cell line from cervix adenocarcinoma) cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab128915 at 2μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1μg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an goat anti-rabbit AlexaFluor®488 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and goat anti-mouse AlexaFluor®594 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.
Flow Cytometry (Intracellular) - Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915)
Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with ab128915 (unpurified) (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 (ab128915, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was goat anti-rabbit Alexa Fluor^® 488 (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (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.
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915)
ab128915 (unpurified), at 1/250, staining GAPDH in MCF7 (human breast adenocarcinoma cell line) cells by immunofluorescence.