Overview

  • Product nameAnti-Bcl-2 antibody [100/D5]
    See all Bcl-2 primary antibodies
  • Description
    Mouse monoclonal [100/D5] to Bcl-2
  • SpecificityThis antibody recognises a protein of 25-26kDa, identified as the bcl-2a oncoprotein. It does not cross-react with bcl-x or bax protein.
  • Tested applicationsFlow Cyt, IHC-P, IHC-Fr, IP, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Cow, Dog, Chinese Hamster

    Does not react with

    Rat
  • Immunogen

    Synthetic peptide corresponding to Bcl-2 aa 41-54.
    Sequence:

    GAAPAPGIFSSQPG-Cys


    Database link: P10415

  • Positive control
    • Follicular lymphomas or tonsil.

Properties

Applications

Our Abpromise guarantee covers the use of ab692 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt 1/10.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB 1/500.
ICC/IF Use a concentration of 5 µg/ml.

Target

  • FunctionSuppresses apoptosis in a variety of cell systems including factor-dependent lymphohematopoietic and neural cells. Regulates cell death by controlling the mitochondrial membrane permeability. Appears to function in a feedback loop system with caspases. Inhibits caspase activity either by preventing the release of cytochrome c from the mitochondria and/or by binding to the apoptosis-activating factor (APAF-1).
  • Tissue specificityExpressed in a variety of tissues.
  • Involvement in diseaseA chromosomal aberration involving BCL2 has been found in chronic lymphatic leukemia. Translocation t(14;18)(q32;q21) with immunoglobulin gene regions. BCL2 mutations found in non-Hodgkin lymphomas carrying the chromosomal translocation could be attributed to the Ig somatic hypermutation mechanism resulting in nucleotide transitions.
  • Sequence similaritiesBelongs to the Bcl-2 family.
  • DomainThe BH4 motif is required for anti-apoptotic activity and for interaction with RAF1 and EGLN3.
  • Post-translational
    modifications
    Phosphorylation/dephosphorylation on Ser-70 regulates anti-apoptotic activity. Growth factor-stimulated phosphorylation on Ser-70 by PKC is required for the anti-apoptosis activity and occurs during the G2/M phase of the cell cycle. In the absence of growth factors, BCL2 appears to be phosphorylated by other protein kinases such as ERKs and stress-activated kinases. Phosphorylated by MAPK8/JNK1 at Thr-69, Ser-70 and Ser-87, wich stimulates starvation-induced autophagy. Dephosphorylated by protein phosphatase 2A (PP2A).
    Proteolytically cleaved by caspases during apoptosis. The cleaved protein, lacking the BH4 motif, has pro-apoptotic activity, causes the release of cytochrome c into the cytosol promoting further caspase activity.
    Monoubiquitinated by PARK2, leading to increase its stability.
  • Cellular localizationMitochondrion outer membrane. Nucleus membrane. Endoplasmic reticulum membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Apoptosis regulator Bcl 2 antibody
    • Apoptosis regulator Bcl-2 antibody
    • Apoptosis regulator Bcl2 antibody
    • AW986256 antibody
    • B cell CLL/lymphoma 2 antibody
    • B cell leukemia/lymphoma 2 antibody
    • B cell lymphoma 2 antibody
    • Bcl 2 antibody
    • Bcl-2 antibody
    • Bcl2 antibody
    • BCL2 protein antibody
    • BCL2_HUMAN antibody
    • C430015F12Rik antibody
    • D630044D05Rik antibody
    • D830018M01Rik antibody
    • Leukemia/lymphoma, B-cell, 2 antibody
    • Oncogene B-cell leukemia 2 antibody
    • PPP1R50 antibody
    • Protein phosphatase 1, regulatory subunit 50 antibody
    see all

Anti-Bcl-2 antibody [100/D5] images

  • ab692 staining Bcl-2 in SK-N-SH cells treated with (R)-(-)-Deprenyl hydrochloride (Selegiline hydrochloride) (ab120604), by ICC/IF. Increase of Bcl-2 expression correlates with increased concentration of (R)-(-)-Deprenyl hydrochloride (Selegiline hydrochloride), as described in literature.
    The cells were incubated at 37°C for 3h in media containing different concentrations of ab120604 ((R)-(-)-Deprenyl hydrochloride (Selegiline hydrochloride)) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab692 (5 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

  • ab692 diluted 1/100 in BSA was incubated with whole cell lysate of human lymphocytes and a Protein G matrix for 12 hours at 4°C to achieve immunoprecipitation.  1,000,000 cells were present in the lysate.

    A modified RIPA buffer was used to lyse cells.

    The IP was washed with PBS/Triton.

    ab692 was used at a 1/1000 dilution for the Western Blot step.

    See Abreview

  • Bcl-2 was localized distinctly in specific cytoplasmic regions in human epithelium (especially in the lower and outmost layers). This image was kindly supplied as part of the review submitted by Marko Nykanen.

     

     

  • Overlay histogram showing Jurkat cells stained with ab692 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab692, 1/10 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

References for Anti-Bcl-2 antibody [100/D5] (ab692)

This product has been referenced in:
  • Sharp AN  et al. Preeclampsia is associated with alterations in the p53-pathway in villous trophoblast. PLoS One 9:e87621 (2014). WB, IHC-P ; Human . Read more (PubMed: 24498154) »
  • Li H  et al. Protective effect of ginsenoside Rg1 on lidocaine-induced apoptosis. Mol Med Rep 9:395-400 (2014). WB ; Human . Read more (PubMed: 24270314) »

See all 21 Publications for this product

Product Wall

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (12)
Sample Mouse Tissue lysate - whole (Heart)
Specification Heart
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Oct 21 2013

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (12)
Sample Rat Tissue lysate - whole (Heart)
Specification Heart
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Oct 14 2013

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (12)
Sample Human Tissue lysate - whole (Heart)
Specification Heart
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Oct 14 2013

Thank you for contacting us. Unfortunately, as we carry over 70,000 products, it simply isn't feasible for us to keep small sample sizes of our products.

However, we are happy to reassure our customers that all of our products are covered by...

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Thank you for your enquiry.

Estimated shelf life of antibodies stored at 4 degrees C is about 1 year, although frequently they last much longer.

I recommend that you follow the storage conditions as noted on the product datasheet. F...

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Thank you for your enquiry. I am sorry to hear you have some concerns regarding the storage of the antibody.

I would suggest to take the antibody out of the freezer and put it at +4oC. The antibody solution does contain some BSA, which can a...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Glioblastoma Initiating Cells (Primary cells))
Loading amount 30 µg
Specification Glioblastoma Initiating Cells (Primary cells)
Gel Running Conditions Reduced Denaturing (12.5)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Nov 16 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Tissue lysate - whole (human islet lysate)
Loading amount 20 µg
Specification human islet lysate
Gel Running Conditions Reduced Denaturing
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Username

Mr. Hao Wu

Verified customer

Submitted Nov 10 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa Cells)
Loading amount 20 µg
Specification HeLa Cells
Gel Running Conditions Non-reduced Denaturing (10)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Aug 23 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (MCF-7)
Loading amount 5 µg
Specification MCF-7
Treatment CDK7 inhibitor
Gel Running Conditions Reduced Denaturing (15)
Blocking step BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Dr. Dean Heathcote

Verified customer

Submitted Jun 12 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"