Anti-Bcl2 Antibody, clone 100

26 kDa

Apoptosis regulator Bcl-2 (UniProt: P10415; also known as Bcl2) is encoded by the BCL2 gene (Gene ID: 596) in human. The members of Bcl-2 family of proteins are known to be involved in either blocking or promoting apoptotic cell death. They act at checkpoints upstream of caspases and share regions of homology termed BH (Bcl Homology) domains. All members of the Bcl-2 family (except Bad and Bid) contain a hydrophobic C-terminus (transmembrane domain), which helps them to attach to membrane. Bcl-2 and other anti-apoptotic members of the Bcl-2 family are located in the outer mitochondrial membrane. Bcl-2 is a single-pass membrane protein that is localized in endoplasmic reticulum, nucleus, and mitochondrial membrane. In mitochondria it is especially enriched at contact sites where the inner and outer membranes come in close proximity. It is reported to counteract the action of pore forming Bax protein. Its transmembrane domain is localized in amino acids 212-233. It contains four BH motifs. BH1 (aa 10-30), BH2 (aa 93-107), and BH4 (aa 136-155) are required for its anti-apoptotic activity. Bcl-2 can be present in cells as homodimer or it can form heterodimers with BAX, BAD, BAK and Bcl-X(L). Heterodimerization with BAX requires intact BH1 and BH2 motifs and is necessary for anti-apoptotic activity. Bcl-2 is shown to suppress apoptosis by controlling the mitochondrial membrane permeability. It functions in a feedback loop system with caspases and is shown to inhibit caspase activity either by preventing the release of cytochrome c from the mitochondria and/or by binding to the apoptosis-activating factor (APAF-1). During apoptosis it is proteolytically cleaved by caspases and the cleaved protein lacking the BH4 motif displays pro-apoptotic activity. Following growth factor stimulation, Bcl-2 can undergo phosphorylation at serine 70 by PKC, which is required for its anti-apoptotic activity.

Epitope: a.a. 41-54

Synthetic peptide corresponding to amino acids 41-54 of human Bcl2 protein. The immunizing sequence is identical in canine and feline.

Flow Cytometry Analysis: A representative lot of this antibody detected Bcl2 in Jurkat cells.
Western Blotting Analysis: 1μg/mL of this antibody detected Bcl2 in Raji cell lysate.
Immunohistochemistry (Paraffin) Analysis: A 1:1,000 dilution of this antibody detected Bcl2 in Human tonsil tissue sections.

Predicted to cross-react with feline and canine, based on sequence homology.

This antibody recognizes Bcl2, Mr 26 kDa.

Format: Purified

Protein G Purified

Purified mouse monoclonal IgG1 in buffer containing 70% storage buffer PBS with 0.1% sodium azide and 30% glycerol. Store at -20ºC.

Stable for 1 year at -20°C from date of receipt.

Handling Recommendations:
Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20ºC. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20ºC may cause glycerol-containing solutions to become frozen during storage.

Control
Tonsil tissue.

Routinely evaluated by western blot on Raji RIPA cell lysate.

Western Blot Analysis:
0.5-2 µg/mL of this lot detected Bcl2 in a Raji RIPA cell lysate.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Replaces: 04-436

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