Westen blot analysis of extracts from various cell lines using BCL2L10 Antibody.
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Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
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
BCL2L10 Antibody detects endogenous levels of total BCL2L10 protein.
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro115 of human BCL2L10. Antibodies are purified by protein A and peptide affinity chromatography.
The Bcl-2 family consists of a number of evolutionarily conserved proteins containing Bcl-2 homology domains (BH) that regulate apoptosis through control of mitochondrial membrane permeability and release of cytochrome c (1-3). Four BH domains have been identified (BH1-4) that mediate protein interactions. The family can be separated into three groups based upon function and sequence homology: pro-survival members include Bcl-2, Bcl-xL, Mcl-1, A1 and Bcl-w; pro-apoptotic proteins include Bax, Bak and Bok; and "BH3 only" proteins Bad, Bik, Bid, Puma, Bim, Bmf, Noxa and Hrk. Interactions between death-promoting and death-suppressing Bcl-2 family members has led to a rheostat model in which the ratio of pro-apoptotic and anti-apoptotic proteins controls cell fate (4). Thus, pro-survival members exert their behavior by binding to and antagonizing death-promoting members. In general, the "BH3-only members" can bind to and antagonize the pro-survival proteins leading to increased apoptosis (5). While some redundancy of this system likely exists, tissue specificity, transcriptional and post-translational regulation of many of these family members can account for distinct physiological roles.
Bcl-2-like 10 (BCL2L10), known as Diva or Boo in mouse (6,7) and Bcl-B in human (8), is a Bcl-2 family member with some unique properties. Expression of the mouse mRNA was detected in multiple embryonic tissues but restricted to adult ovary and testis (6,7); human Bcl-B appears to be more widely expressed (8). BCL2L10 contains BH1, 2, and 4 domains as well as a putative carboxy-terminal transmembrane domain. While some studies report the presence of a pro-apoptotic BH3 domain in BCL2L10, conflicting reports indicate an incomplete or absent BH3 domain (7-9). Similarly, some studies indicate that BCL2L10 induces apoptosis (6,9) while other data implies a role in suppressing cell death (7,8,10). BCL2L10 may function by differentially binding other Bcl-2 family members and through interaction with the apoptosome protein Apaf-1 (6,7). Despite its restricted expression in mice, Diva knockouts were fertile and exhibit no obvious developmental defects (11).
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