|Reactivity||Human, Mouse, Rat|
|Product Name||Anti-GAPDH Rabbit Monoclonal Antibody, HRP Conjugated|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time. Avoid repeated freezing and thawing.|
|Description||Anti-GAPDH Rabbit Monoclonal Antibody, HRP Conjugated tested for WB in Human, Mouse, Rat|
|Cite This Product||Anti-GAPDH Rabbit Monoclonal Antibody, HRP Conjugated (Boster Biological Technology, Pleasanton CA, USA, Catalog # H00227)|
|Immunogen||A synthesized peptide derived from human GAPDH(HRP conjugated)|
|Reactivity||Human, Mouse, Rat|
Assay Dilutions Overview
Images And Assay Conditions
Western blot analysis of GAPDH expression in (1) Jurkat cell lysate; (2) A375 cell lysate; (3) Human hippocampus lysate; (4) Human fetal liver lysate; (5) COS-1 cell lysate; (6) Raw2647 cell lysate; (7) Mouse kidney lysate; (8) PC-12 cell lysate; (9) Rat brain lysate with GAPDH Antibody (H00227).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-GAPDH monoclonal antibody (Catalog # H00227) overnight at 4℃, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for GAPDH
Protein Target Info (Source: Uniprot.org)
|Protein Name||Glyceraldehyde-3-phosphate dehydrogenase|
|Alternative Names||Glyceraldehyde-3-phosphate dehydrogenase;GAPDH;220.127.116.11;Peptidyl-cysteine S-nitrosylase GAPDH;2.6.99.-;GAPDH;GAPD;CDABP0047, OK/SW-cl.12;|
|Subcellular Localization||Cytoplasm, cytosol . Nucleus . Cytoplasm, perinuclear region . Membrane . Cytoplasm, cytoskeleton . Translocates to the nucleus following S- nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions. .|
|Molecular Weight||36053 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein Function||Has 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. Modulates the organization and assembly of the cytoskeleton. Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules (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. Component of the GAIT (gamma interferon- activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes. 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. .|
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