Uses
Monoclonal Anti-GAPDH antibody produced in mouse is suitable for western blotting using:
- protein extracted from heart tissue of mice at a working dilution of 1:25,000
- myelin and axogliasomal fractions from human CNS
- nuclear and cytoplasmic fractions from TBP-13Q and TBP-105Q PC12 cells following recovery from heat shock
- protein from bovine immortalized luteal endothelial cells
- renal tubular epithelial cell extract
- proteins from mouse embryonic fibroblasts
- protein extract from ventricular myocardium tissues
- A431 total cell extract at a working concentration of 0.025-0.05μg/mL
It is also suitable for immunostaining using leiomyomas and leiomyosarcomas. The antibody can also be used for immunocytochemistry, indirect ELISA and microarray.
General Description
Monoclonal Anti-GAPDH (mouse IgM isotype) is derived from the hybridoma GAPDH-71.1 produced by the fusion of mouse myeloma cells (NS1 cells) and splenocytes from BALB/c mice immunized with rabbit GAPDH (glyceraldehyde-3-phosphate dehydrogenase). The gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is mapped to human chromosome 12p13. The protein localizes in the cytoplasm but can be translocated to the nucleus depending on cellular conditions.
Biochem/physiol Actions
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a tetramer containing identical chains. It catalyzes the reversible oxidative phosphorylation of glyceraldehyde-phosphate, which is a critical energy-yielding step in carbohydrate metabolism. It binds to several proteins including actin, tubulin, amyloid precursor, polyglutamine peptides, DRPLA (dentatorubral-pallidoluysian atrophy), and huntingtin. Protein kinase Cι/λbinds and phosphorylates GAPDH. Phosphorylated GAPDH associates with cytoskeletal elements and controls microtubule dynamics in the early secretory pathway. Poly(ADP-ribose) polymerase-1 (PARP1) interacts with GAPDH and thereby mediates brain damage in the presence of oxidative/nitrosative stress. GAPDH forms a part of OCA-S, the multicomponent OCT1 (octamer-motif-binding factor) coactivator complex, which is involved in the S phase-dependent histone H2B transcription. This association is responsible for linking H2B transcriptional machinery to cell cycle regulation and to the cellular metabolic state. GAPDH is also a component of the functional GAIT (interferon-γ-activated inhibitor of translation) mRNP (messenger ribonucleoprotein). GAPDH expression is dysregulated during melanoma progression.
