Product nameAnti-GAPDH antibody [3E8AD9] - Loading Control (HRP)
See all GAPDH primary antibodies
DescriptionMouse monoclonal [3E8AD9] to GAPDH - Loading Control (HRP)
Tested applicationsSuitable for: WB, IHC-Pmore details
Species reactivityReacts with: Human
Tissue, cells or virus corresponding to Human GAPDH. Purified protein from Human erythrocytes.
- WB: HepG2 whole cell lysate. IHC-P: FFPE human liver (normal) tissue sections.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at +4°C. Stable for 12 months at -20°C. Store In the Dark.
Storage bufferpH: 7.4
Preservative: 0.1% Proclin
Constituents: 30% Glycerol, 1% BSA, PBS
Concentration information loading...
Purification notesPurity >95% by SDS-PAGE.
Light chain typekappa
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Energy Metabolism
Our Abpromise guarantee covers the use of ab198306 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/5000. Detects a band of approximately 36 kDa (predicted molecular weight: 36 kDa).|
|IHC-P||1/50. 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 liver tissue*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins, and incubated overnight at +4°C with ab198306 at 1/50 dilution. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature. The section was 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
Anti-GAPDH antibody [3E8AD9] - Loading Control (HRP) (ab198306) at 1/5000 dilution + HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 36 kDa
Observed band size: 36 kDa
Exposure time: 2 minutes
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 ab198306 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.
ab198306 has not yet been referenced specifically in any publications.