Anti-Bif-1 (Bax interacting factor-1, SH3GLB1, Bif-1, CGI-61, KIAA0491, SH3-domain GRB2-like endophi

The Bcl-2 family consists of anti-apoptotic (or cell survival) genes, such as Bcl-2, Bcl-XL and pro-apoptotic (or cell death) genes including Bax and BAD. In addition to its anti-apoptotic effect, Bcl-2 also inhibits progression through the cell cycle. Functional interactions between family members as well as binding to other cellular proteins modulate their activities. Bcl-2 family members are important in normal tissue development, homeostasis, and disease states. Bcl-2 prevents cell death by forming heterodimers with Bax and Bad (1). Bax promotes cell death by forming homodimers, which can be blocked by forming heterodimers with Bcl-2 or BCL-XL. Recently, using yeast two-hybrid system Cuddeback et al (2) have identified a cDNA encoding a 365 amino acid protein, which interacts physically with Bax protein and influences cell life and death. Bif-1 contains a Src homology 3 (SH3) domain in the C-terminus. It does not contain any of the conserved Bcl-2 homology (BH) domains of the Bcl-2 family proteins. Human Bif-1 protein shares 96% and 42% identity at the amino acids level with mouse and C. elegans, respectively. Overexpression of Bif-1 in transfected cell lines promotes Bax conformational change, caspase activation, and apoptotic cell death following growth factor withdrawal. Bif-1 RNA is abundantly expressed in heart, skeletal muscle, kidney, and placenta. Domain Information: BAR domain, Src homology 3 domains , Gene Ontology Terms: , Application(s): Peptide ELISA: antibody detection limit dilution 1:32,000. Western Blot: Approx 38kD band observed in lysates of Human T cell line Jurkat and in Human Lung and Placenta (calculated MW of 40.8kD according to NP_057093.1). Recommended concentration: 0.3-1.0 ug/ml. A minor band of unknown identity was also consistently observed at 50kD in Jurkat and Placenta. This band was successfully blocked by incubation with the immunizing peptide. We would appreciate any feedback from people in the field-have any results been reported with other antibodies/lysates? Have any further splice variants/modified forms been reported?, Staining (0.3ug/ml) of Jurkat lysate (RIPA buffer, 35ug total protein per lane). Primary incubated for 1 hour. Detected by western blot using chemiluminescence. Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. Peptide Blocking: Corresponding peptide is available for Peptide Blocking studies. See B1098-01B-P. Antibody is typically 0.5mg/ml and peptide is supplied as a 100ul pellet. When peptide is reconstituted in 200ul water, the concentration would also be 0.5mg/ml. To start, the best ratio would be 1:1 (molar excess of peptide relative to antibody when identical volumes are mixed). Mix equal volumes of peptide and antibody at the required dilution and leave at ambient temperature. It is best is to have two identical blots to be incubated with equal amount of antibody, but one with the antibody pre-adsorbed to the peptide for 20min. Then incubate and develop the two blots in parallel.