Detects human Bcl-2 in direct ELISAs and Western blots. In direct ELISAs, approximately 25‑40% cross‑reactivity with recombinant mouse (rm) Bcl‑2 is observed and no cross-reactivity with recombinant human BCL2L12 is observed. In Western blots, 100% cross-reactivity with rmBcl-2 is observed.
Monoclonal Mouse IgG2B Clone # 625509
Protein A or G purified from hybridoma culture supernatant
E. coli-derived recombinant human Bcl-2 Ala2-Asp211 Accession # P10415
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Detection of Human, Mouse, and Rat Bcl‑2 by Western Blot.
Western blot shows lysates of THP‑1 human acute monocytic leukemia cell line, KG‑1 human acute myelogenous leukemia cell line, CTLL‑2 mouse cytotoxic T cell line, and NRK rat normal kidney cell line. PVDF Membrane was probed with 0.1 µg/mL of Mouse Anti-Human/Mouse/Rat Bcl‑2 Monoclonal Antibody (Catalog # MAB8272) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Bcl‑2 at approximately 24 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Bcl‑2 in Human Lymphoma.
Bcl‑2 was detected in immersion fixed paraffin-embedded sections of human lymphoma using Mouse Anti-Human/Mouse/Rat Bcl‑2 Monoclonal Antibody (Catalog # MAB8272) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Bcl-2 is a member of a family of proteins that regulates outer mitochondrial membrane permeability (1, 2). Bcl-2 is an anti-apoptotic member that prevents release of cytochrome c from the mitochondria intermembrane space into the cytosol. Bcl-2 is present on the outer mitochondrial membrane and is also found on other membranes in some cell types. Natural Bcl-2 contains a carboxyl-terminal mitochondria targeting sequence. Recombinant Bcl-2, missing the mitochondrial targeting sequence, maintains its ability to neutralize pro-apoptotic Bcl-2 family members. Neutralization by Bcl-2 appears to be through binding the BH3 region of pro-apoptotic Bcl-2 family members. This activity does not require the mitochondrial targeting sequence.
Gross, A. et al. (1999) Genes and Develop. 13:1899.
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