GAPDH (glyceraldehyde-3-phosphate dehydrogenase) is an intracellular homotetramerizing enzyme, which catalyzes oxidation of glyceraldehyde-3-phosphate into 1,3-bisphosphoglycerate during glycolysis, and has also functions in DNA replication, gene expression, and microtubule bundling. As it binds to many proteins, its functions are very broad. GAPDH plays also roles in cancer progression, apoptosis, Huntington disease and Alzheimer´s disease. For its ubiquitous expression this antigen belongs to those that serve as loading controls in Western blotting experiments. However, one must keep in mind that under hypoxic conditions GAPDH level can be significantly (or even strongly) increased, as well as under the effect of insulin, nitric oxide or p53.
The mouse monoclonal antibody FF26A recognizes human GAPDH and can be used in Western blotting (both reducing and non-reducing conditions), immunohistochemistry (frozen sections), and immunoprecipitation application.
Currently available format:
Western blotting analysis was performed on whole cell extracts (RIPA lysis buffer) of HeLa, HEK 293T, Jurkat and Raji cell lines mixed and heated (100°C, 5 min) with reducing (2-mercaptoethanol) or non-reducing SDS-loading buffer. Samples were resolved using 15% Tris-glycine SDS gel electrophoresis. Nitrocellulose membrane blot was probed simultaneously with mouse IgM monoclonal antibody TU-10 (1 µg/ml) and anti-GAPDH mouse IgG1 monoclonal antibody FF26A (1 µg/ml). Subclass-specific secondary antibodies IRDye 680RD Goat-anti-Mouse IgM (red) and IRDye 800CW Goat-anti-Mouse IgG (green) were used for multiplex fluorescent Western blot detection.
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