BH3-Only Protein, Antibody Sampler Set, BioAssay(TM)

Apoptosis plays a major role in normal organism development, tissue homeostasis, and removal of damaged cells. This process is regulated by the interplay of pro- and anti-apoptotic members of the B-cell lymphoma 2 (Bcl-2) family. The BH3-only (Bcl-2-homology domain 3 only) proteins are a pro-apoptotic subgroup of the Bcl-2 family that are critical initiators of lymphocyte apoptosis and other processes. Unlike the other members of the Bcl-2 family which contain at least two Bcl-2 homology (BH) domains, these proteins contain just one, the BH3 domain. Some of the members of this family, such as Hrk, may show little sequence homology to the others, but all BH-only proteins can bind to at least some pro-survival Bcl- 2-like proteins through this BH3 domain and trigger apoptosis when over-expressed. For example, the proteins Bad, Bim and Bmf regulate apoptosis by forming heterodimers with the anti-apoptotic proteins Bcl-2 and Bcl-xL. Bim, Noxa, and PUMA are activated by p53 following DNA damage although the exact mechanisms are not entirely clear. Noxa may also be involved with the activation of Bik, a BH3-only protein whose localization in the endoplasmic reticulum (ER) suggests that Bik may play a role in ER- stress-induced apoptosis. While full-length Bid also promotes apoptosis when expressed, its activity is enhanced following cleavage by enzymes such as capsase-8, suggesting that Bid may play a key role in death-receptor induced apoptosis. Rabbit polyclonal antibodies were raised against peptide sequences corresponding to each of the target proteins. Specificity: These polyclonal antibodies can be used for detection of Bad, Bid, Bik, Bim, Bmf, Hrk, Noxa and PUMA by immunoblot at 1-2ug/ml and their respective proteins via immunohistochemistry at 1-10ug/ml. Kit Components: Bad (Rb x Hu): 1x25ug Bid, CT (Rb x Hu): 1x25ug Bik (Rb x Hu): 1x25ug Bim, ID (Rb x Hu): 1x25ug Bmf, NT (Rb x Hu): 1x25ug Hrk, CT (Rb x Hu): 1x25ug Noxa (Rb x Hu): 1x25ug PUMA, CT (Rb x Hu): 1x25ug Applications: Suitable for use in Western Blot and Immunoblot. Other applications not tested. Recommended Dilution: Immunoblot: 1-2ug/ml Immunohistochemistry: 1-10ug/ml Optimal dilutions to be determined by the researcher. Positive Control: Anti-Bim: K562 Cell Lysate Anti-PUMA: K562 Cell Lysate Storage and Stability: May be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.