Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Simple Western, Immunoprecipitation

Cited:

Immunohistochemistry-Paraffin, Western Blot, Bioassay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Bcl-2 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant mouse Bcl-2

Specificity

Detects human and mouse Bcl-2 in Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human/Mouse Bcl‑2 Antibody

Detection of Human/Mouse Bcl-2 antibody by Western Blot.

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.

Bcl-2 antibody in Human Breast Cancer Tissue by Immunohistochemistry (IHC-P).

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 of Mouse Bcl‑2.

Immunoprecipitation of Mouse Bcl‑2.

Bcl‑2 was immunoprecipitated from lysates (3 x 106cells) 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 antibody by Simple WesternTM.

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.

Detection of Bcl-2 by Western Blot

Detection of Bcl-2 by Western Blot

Glycyrrhizic acid induces the apoptosis of colorectal cancer cells by inhibiting SIRT3 in vitro. (a,b) 0 or 20 μmol/L glycyrrhizin stimulated colorectal cancer cells for 24 hours, flow cytometry was used to detect the apoptosis of colorectal cancer cells (a) and statistical comparison (b). (c) Western blot analysis of Bax and Bcl2 protein expression in colorectal cancer cells after stimulating with 0 or 20 μmol/L glycyrrhizic acid for 24 hours. (d) Elisa kit was used to determine the activity of caspase 3 in colorectal cancer cells after stimulating with 0 or 20 μmol/L glycyrrhizic acid for 24 hours. Each test is repeated at least 3 times independently. *** P < 0.001 vs WT group, and ### P < 0.001 vs 20 μM GA group. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/34747319), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse Bcl‑2 Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human breast cancer tissue

Immunoprecipitation

1 µg/106 cells
Sample: M‑NFS‑60 mouse myelogenous luekemia lymphoblast cell line, see our available Western blot detection antibodies

Simple Western

50 µg/mL
Sample: KG‑1 human acute myelogenous leukemia cell line

Western Blot

1 µg/mL
Sample: KG‑1 human myeloid leukemia cell line and M‑NFS‑60 mouse myelogenous leukemia lymphoblast cell line

Reviewed Applications

Read 2 reviews rated 5 using AF810 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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

Alternate Names

Bcl2

Entrez Gene IDs

596 (Human); 12043 (Mouse); 24224 (Rat)

Gene Symbol

BCL2

Additional Bcl-2 Products

Product Documents for Human/Mouse Bcl‑2 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse Bcl‑2 Antibody

For research use only

Citations for Human/Mouse Bcl‑2 Antibody

Customer Reviews for Human/Mouse Bcl‑2 Antibody (2)

5 out of 5
2 Customer Ratings
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  • Human/Mouse Bcl-2 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Brain (glial cell in hippocampus)
    Species: Human and Mouse
    Verified Customer | Posted 06/17/2024
    The conjugation of R&D Systems' Bcl-2 antibody for immunofluorescence has been pivotal in advancing our research on apoptosis regulation. Bcl-2 protein's role in cell survival necessitates accurate detection, making the choice of antibody and conjugation method critical. We selected R&D Systems' antibody for its established reliability and effectiveness in previous studies. Conjugating the antibody to fluorescent dyes enabled precise localization and visualization of Bcl-2 expression in various cellular contexts. The conjugation process was straightforward, and the resulting fluorescence signals were robust and specific, providing clear insights into protein distribution and dynamics within cells. Validation involved using appropriate controls and optimizing staining protocols to maximize signal-to-noise ratios. The antibody consistently demonstrated high specificity, effectively distinguishing Bcl-2 from non-specific background fluorescence. In conclusion, R&D Systems' Bcl-2 antibody conjugated for immunofluorescence has been instrumental in our research, offering reliable and detailed visualization of apoptotic mechanisms. Its performance in conjugation and subsequent imaging has enhanced our understanding of Bcl-2's cellular functions, making it a valuable tool for both basic research and potential therapeutic applications
    Human/Mouse Bcl‑2 Antibody AF810
  • Human/Mouse Bcl-2 Antibody
    Name: Jothi Jayavel
    Application: Western Blot
    Sample Tested: MDA-MBA-231 human breast cancer cell line
    Species: Human
    Verified Customer | Posted 01/11/2018
    Human/Mouse Bcl‑2 Antibody AF810

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Protocols

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