Anti-Human Bag-1

Human Bag-1

This monoclonal antibody has been 0.2 µm filtered and lyophilized from modified Dulbecco's phosphate buffered saline (1X PBS) pH 7.2 - 7.3 containing 5.0% w/v trehalose with no calcium, magnesium or preservatives present.

The lyophilized antibody can be stored desiccated at -20°C to -70°C for up to twelve months. The reconstituted antibody can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted antibody, aseptically aliquot into working volumes and store at -20°C to -70°C in a manual defrost freezer. Avoid repeated freeze thaw cycles. No detectable loss of activity was observed after six months.

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Immunoprecipitation: Anti-human Bag-1 (3 μg) is added to 1 mL of cell extract and the mixture is rocked for 1 - 16 hours at 4° C. Rabbit anti-mouse IgG (6 μg) is then added and after 30 minutes, Immunoprecipitin (50 μL of a 20% suspension) is then added and the mixture is rocked in the cold for an additional 30 minutes. The Immunoprecipitin-absorbed complexes are centrifuged for 30 seconds in an Eppendorf centrifuge, resuspended in wash buffer by trituration with a glass Pastuer pipet, and then repelleted. The complexes are washed a total of three times with extraction buffer, and then suspended in phosphate buffered saline and transferred to a new tube before the final centrifugation. The washed pellet is suspended in 25 - 50 μL of 2x SDS gel sample buffer (20 mM dithiothreitol, 6% SDS, 0.25 M Tris, pH 6.8, 10% glycerol, and bromophenyl blue) by vortexing and then incubated for 3 minutes in a boiling water bath. Immunprecipitin is pelleted and the supernatant is loaded on a polyacrylamide gel.

Mouse Anti-Human Bag-1 (Clone CUV02) recognizes an epitope on Human Bag-1. This monoclonal antibody was purified using multi-step affinity chromatography methods such as Protein A or G depending on the species and isotype.

Bcl-2-associated athanogene 1 (BAG-1) was first identified as an anti-apoptotic bcl-2-binding protein. Later it was found to bind the molecular chaperones Hsp70 and Hsc70 through its carboxy-terminal sequence (termed the Bag domain), resulting in the inhibition of the refolding activity of these chaperones. It is thought that by binding and inhibiting these molecular chaperones, BAG-1is able to modulate the expression level of proteins requiring chaperones to fold correctly. One such group of proteins that are affected is glucocorticoid receptors. Other reports have suggested that the level of BAG-1 expression correlates with the aggressiveness of various cancers. Multiple isoforms of BAG-1 are known to exist.