The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 0.5 - 1 µg/ml. Predicted molecular weight: 36 kDa.
Use at an assay dependent concentration.
Use a concentration of 1 µg/ml.
功能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.
通路Carbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5.
序列相似性Belongs to the glyceraldehyde-3-phosphate dehydrogenase family.
翻译后修饰S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus. ISGylated.
细胞定位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.
ICC/IF image of ab83957 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat 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 ab83957 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- chicken (ab96947) IgY (H+L) used at a 1/250 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.
Western blot - GAPDH antibody (ab83957)
Lane 1 : Anti-GAPDH antibody (ab83957) at 1 µg/ml Lane 2 : Anti-GAPDH antibody (ab83957) at 2 µg/ml
Lane 1 : HeLa cell lysate Lane 2 : HeLa cell lysate