Anti-Bcl-2 [100/D5] antibody (ab692)
- Product nameAnti-Bcl-2 [100/D5] antibodySee all Bcl-2 primary antibodies ...
- DescriptionMouse 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/IF more details
- Species reactivityReacts with: Mouse, Human
Predicted to work with: Cow, Dog, Chinese Hamster
Does not react withRat
Synthetic peptide corresponding to Bcl-2 aa 41-54.
- Positive control
- Follicular lymphomas or tonsil.
- Storage instructionsStore at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
- Storage bufferPreservative: 0.09% Sodium Azide
Constituents: 1% BSA, Tissue culture supernatant, PBS, pH 7.3
- Concentration information loading...
- PurityImmunogen affinity purified
- Clonality Monoclonal
- Clone number100/D5
- Light chain typekappa
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.
|Flow Cyt||Flow Cyt: 1/10.|
|IHC-P||IHC-P: Use at an assay dependent concentration.|
|IHC-Fr||IHC-Fr: Use at an assay dependent concentration.|
|IP||IP: Use at an assay dependent concentration.|
|ICC/IF||ICC/IF: Use a concentration of 5 µg/ml.|
- 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.
modificationsPhosphorylation/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.
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Anti-Bcl-2 [100/D5] antibody 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.
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 [100/D5] antibody (ab692)
This product has been referenced in:
- Jiao LR et al. MicroRNAs targeting oncogenes are down-regulated in pancreatic malignant transformation from benign tumors. PLoS One 7:e32068 (2012). WB ; Human . Read more (PubMed: 22384141) »
- Melarangi T et al. Glucocorticoid resistance in chronic lymphocytic leukaemia is associated with a failure of upregulated Bim/Bcl-2 complexes to activate Bax and Bak. Cell Death Dis 3:e372 (2012). WB . Read more (PubMed: 22898870) »