Product nameAnti-GAPDH antibody [EPR6256] - Loading Control (Biotin)
See all GAPDH primary antibodies
DescriptionRabbit monoclonal [EPR6256] to GAPDH - Loading Control (Biotin)
Tested applicationsSuitable for: WB, IHC-Pmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse
Synthetic peptide within Human GAPDH aa 250-350. The exact sequence is proprietary.
Database link: P04406
- WB: HEK293, HeLa, MCF7 and SH SY5Y whole cell lysates. IHC-P: FFPE human testis (normal) tissue sections.
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. Store In the Dark.
Storage bufferpH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 30% Glycerol, 1% BSA, PBS
Concentration information loading...
PurityProtein A purified
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Energy Metabolism
- Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915)
- Anti-GAPDH antibody [EPR6256] - Loading Control (HRP) (ab185059)
- Anti-GAPDH antibody [EPR6256] - BSA and Azide free (ab186930)
- Anti-GAPDH antibody [EPR6256] (Alexa Fluor® 647) (ab215227)
- Anti-GAPDH antibody [EPR6256] (PerCP) (ab216377)
- Anti-GAPDH antibody [EPR6256] (Allophycocyanin) (ab221270)
- HeLa nuclear extract lysate (ab14655)
- HepG2 cytoplasmic extract lysate (ab14659)
- HepG2 nuclear extract lysate (ab14660)
- MCF7 nuclear extract lysate (ab14860)
- HeLa whole cell lysate (ab150035)
- MCF7 membrane extract lysate (ab29539)
- HeLa membrane extract lysate (ab29547)
- HEK293 whole cell lysate (ab7902)
Our Abpromise guarantee covers the use of ab195904 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. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).|
|IHC-P||1/150. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
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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IHC image of GAPDH staining in a section of formalin-fixed paraffin-embedded normal human testis*, performed on a Leica BondTM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins, before blocking of endogenous biotin using ab64212. The section was then incubated with ab195904, 1/150 dilution, for 15 mins at room temperature and detected using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.
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
All lanes : Anti-GAPDH antibody [EPR6256] - Loading Control (Biotin) (ab195904) at 1/10000 dilution
Lane 1 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 2 :
HeLa whole cell lysate (ab150035)
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 4 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 36 kDa
Observed band size: 36 kDa
Exposure time: 2 seconds
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab195904 overnight at 4°C. Antibody binding to the target was detected using a HRP conjugated streptavidin amplification step, and visualised using ECL development solution ab133406.
ab195904 has not yet been referenced specifically in any publications.