|MW (kDa)||19, 21|
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: Loading of prestained molecular weight markers (#59329, 10 µl/lane) to verify electrotransfer and biotinylated protein ladder (#7727, 10 µl/lane) to determine molecular weights are recommended.
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
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues within a central region of Bit1. Antibodies were purified by protein A and peptide affinity chromatography.
Bit1 is a mitochondrial protein discovered as a regulator of anoikis, a process of apoptosis triggered by the loss of cell adhesion to the extracellular matrix (1,2). Like several other mitochondrial apoptotic regulators (Cytochrome c, Smac, HtrA2, AIF, EndoG), Bit1 is released into the cytoplasm during apoptosis (1,3). Cytoplasmic Bit1 associates with the Groucho family of transcriptional regulator AES (1,4). Interaction between Bit1 and AES promotes apoptosis, and this complex is suppressed by integrin mediated cell attachment.
UniProt ID: Q9Y3E5
Entrez-Gene Id: 51651
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