The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Detects a band of approximately 37 kDa (predicted molecular weight: 35.8 kDa).
Use a concentration of 1 µg/ml.
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 (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.
Carbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5.
Belongs to the glyceraldehyde-3-phosphate dehydrogenase family.
S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus. ISGylated.
Cytoplasm > 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.
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody - Loading Control (ab9483)
ICC/IF image of ab9483 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal donkey serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9483, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 donkey anti-goat IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa cells at 1µg/ml.
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody - Loading Control (ab9483)This image is courtesy of an Abreview submitted by Kirk McManus
ab9483 staining GAPDH in HeLa by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehydeand. Samples were incubated with primary antibody (1/200) for 30 minutes. An undiluted Alexa Fluor® 568-conjugated Donkey anti-goat polyclonal was used as the secondary antibody.
Foley B et al. Editor's Highlight: Screening ToxCast Prioritized Chemicals for PPARG Function in a Human Adipose-Derived Stem Cell Model of Adipogenesis. Toxicol Sci155:85-100 (2017).
Read more (PubMed: 27664422) »
Wei D et al. Genistein suppresses retinoblastoma cell viability and growth and induces apoptosis by upregulating miR-145 and inhibiting its target ABCE1. Mol Vis23:385-394 (2017).
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