Product nameAnti-GAPDH antibody [EPR6256] - Loading Control
See all GAPDH primary antibodies
DescriptionRabbit monoclonal [EPR6256] to GAPDH - Loading Control
Tested applicationsSuitable for: WB, IP, Flow Cyt, ICC/IF, IHC-Pmore details
Species reactivityReacts with: Human, African green monkey
Does not react with: Mouse
Synthetic peptide within Human GAPDH aa 250 to the C-terminus. The exact sequence is proprietary.
Database link: P04406
- WB: 293T, HeLa, HepG2, HUVEC, MCF7 and SH-SY5Y cell lysates ICC/IF: HeLa and MCF7 cells. IHC-P: Human pancreas tissue. Flow Cyt: HeLa cells.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
PurityTissue culture supernatant
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Energy Metabolism
- Anti-GAPDH antibody [EPR6256] - Loading Control (HRP) (ab185059)
- Anti-GAPDH antibody [EPR6256] - BSA and Azide free (ab186930)
- Anti-GAPDH antibody [EPR6256] - Loading Control (Biotin) (ab195904)
- Anti-GAPDH antibody [EPR6256] (Phycoerythrin) (ab209929)
- Anti-GAPDH antibody [EPR6256] (Phycoerythrin) (ab214521)
- Anti-GAPDH antibody [EPR6256] (Alexa Fluor® 647) (ab215227)
- Anti-GAPDH antibody [EPR6256] (PerCP) (ab216377)
- Anti-GAPDH antibody [EPR6256] (Allophycocyanin) (ab221270)
Our Abpromise guarantee covers the use of ab128915 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/10000 - 1/50000. Detects a band of approximately 35 kDa (predicted molecular weight: 36 kDa).|
|IP||1/10 - 1/100.|
|Flow Cyt||1/100 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|ICC/IF||1/250 - 1/500.
FunctionHas both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. 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). 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.
PathwayCarbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5.
Sequence similaritiesBelongs to the glyceraldehyde-3-phosphate dehydrogenase family.
modificationsS-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
Cellular localizationCytoplasm > cytosol. Nucleus. Cytoplasm > perinuclear region. Membrane. Translocates to the nucleus following S-nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions.
- Information by UniProt
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All lanes : 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
Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 3 : HepG2 (human liver hepatocellular carcinoma cell line) cell lysate
Lane 4 : HUVEC (human umbilical vein endothelial cell line) cell lysate
Lane 5 : MCF7 (human breast adenocarcinoma cell line) cell lysate
Lane 6 : SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate
Lysates/proteins at 10 µg per lane.
All lanes : HRP labelled Goat anti-Rabbit IgG at 1/2000 dilution
Predicted band size: 36 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?
IHC image of ab128915 staining GAPDH in human pancreas* formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab128915, 1:250 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
ab128915 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 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 (ab150081) at 2 μg/ml (shown in green) and goat anti-mouse AlexaFluor®594 (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.
Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with ab128915 (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) (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.
ab128915 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.
ab128915, at 1/250, staining GAPDH in MCF7 (human breast adenocarcinoma cell line) cells by immunofluorescence.
This product has been referenced in:
- Chen W et al. Gemcitabine combined with an engineered oncolytic vaccinia virus exhibits a synergistic suppressive effect on the tumor growth of pancreatic cancer. Oncol Rep 41:67-76 (2019). Read more (PubMed: 30365143) »
- Qin N et al. Deubiquitinating enzyme 4 facilitates chemoresistance in glioblastoma by inhibiting P53 activity. Oncol Lett 17:958-964 (2019). Read more (PubMed: 30655854) »