Human/Mouse Bcl-2 Antibody

(3 citations)   
  • Species Reactivity
    Human, Mouse
  • Specificity
    Detects human and mouse Bcl-2 in Western blots.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant mouse Bcl-2
  • Formulation
    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.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Simple Western
    50 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Immunoprecipitation
    1 µg/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human/Mouse Bcl‑2 by Western Blot. Western blot shows lysates of KG‑1 human myeloid leukemia cell line and M‑NFS‑60 mouse myelogenous leukemia lymphoblast cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human/Mouse Bcl‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF810) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Bcl‑2, at approximately 25 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Immunohistochemistry
Bcl-2 in Human Breast Cancer Tissue. Bcl‑2 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using 15 µg/mL Goat Anti-Human/
Mouse Bcl‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF810) overnight at 4 °C. Tissue was stained (red) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Immunoprecipitation
Immunoprecipitation of Mouse Bcl‑2. Bcl‑2 was immunoprecipitated from lysates (3 x 106 cells) of M‑NFS‑60 mouse myelogenous leukemia lymphoblast cell line following incubation with 3 µg Goat Anti-Human/Mouse Bcl‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF810) for 1 hour at 4 °C. Bcl‑2-antibody complexes were absorbed using Protein G (Sigma). Immunoprecipitated Bcl‑2 was detected by Western blot using 1 µg/mL Goat Anti-Human/Mouse Bcl‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF810). View our recommended buffer recipes for immunoprecipitation.
Detection of Human Bcl‑2 by Simple WesternTM. Simple Western lane view shows lysates of KG‑1 human acute myelogenous leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for Bcl‑2 at approximately 33 kDa (as indicated) using 50 µg/mL of Goat Anti-Human/Mouse Bcl‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF810) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    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.
Background: Bcl-2

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.

 

  • References:
    1. Gross, A. et al. (1999) Genes and Develop. 13:1899.
    2. Kroemer, G. (1997) Nature Med. 3:614.
  • Long Name:
    B Cell Lymphoma/Leukemia 2
  • Entrez Gene IDs:
    596 (Human); 12043 (Mouse); 24224 (Rat)
  • Alternate Names:
    apoptosis regulator Bcl-2; B-cell CLL/lymphoma 2; Bcl2; Bcl-2
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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Species
Applications
Sample Type
  1. TCDD promotes lung tumors via attenuation of apoptosis through activation of the Akt and ERK1/2 signaling pathways.
    Authors: Chen R, Siao S, Hsu C, Chang C, Chang L, Wu C, Lin P, Wang Y
    PLoS ONE, 2014;9(6):e99586.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: Bioassay
  2. Molecular profiling of cervical cancer progression.
    Authors: Hagemann T, Bozanovic T, Hooper S, Ljubic A, Slettenaar VI, Wilson JL, Singh N, Gayther SA, Shepherd JH, Van Trappen PO
    Br. J. Cancer, 2007;96(2):321-8.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
  3. The anti-apoptotic effect of Notch-1 requires p56lck-dependent, Akt/PKB-mediated signaling in T cells.
    Authors: Sade H, Krishna S, Sarin A
    J. Biol. Chem., 2004;279(4):2937-44.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
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