For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu523 of human ATP5B protein.
ATP5B (ATP5F1B) is the catalytic β subunit of the F1 portion of the F1F0 ATP synthase, which is responsible for the majority of ATP synthesis in oxidative phosphorylation (1). ATP5B is present in the inner membrane of mitochondria in all types of cells, where it functions as an ATP provider and plays an important role in maintaining intracellular energy homeostasis. A high level of ATP5B is associated with cancer proliferation, metastasis, and drug resistance (2-4). An increased level of ATP5B plays an important protective role in decreasing renal fibrosis (5). Cell surface expression of ATP5B has been found in various cell types, including tumor, endothelial, and neuronal cells (6), where it is involved in multiple biological processes depending on the cell types. These include promoting viral entry (7), serving as a ligand for tumor-T cell recognition (8), generating extracellular ATP, contributing to intracellular PH homeostasis (9), and facilitating tumor metastasis (10). Inhibition of ATP5B has been proposed as a potential treatment for aggressive cancer (11,12).
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