The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. Detects a band of approximately 26 kDa (predicted molecular weight: 26 kDa).
Use a concentration of 1 - 5 µg/ml.
Use at an assay dependent concentration.
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). May attenuate inflammation by impairing NLRP1-inflammasome activation, hence CASP1 activation and IL1B release (PubMed:17418785).
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.
DomainBH1 and BH2 domains are required for the interaction with BAX and for anti-apoptotic activity. The BH4 motif is required for anti-apoptotic activity and for interaction with RAF1 and EGLN3. The loop between motifs BH4 and BH3 is required for the interaction with NLRP1.
Post-translational 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. Ubiquitinated by SCF(FBXO10), leading to its degradation by the proteasome.
Predicted band size : 26 kDa Observed band size : 26 kDa
Anti-Bcl-2 antibody (ab47489)
Immunohistochemical analysis of paraffin-embedded breast carcinoma, using ab47489 at a 1/50 dilution.
Left image: Untreated
Right image: Treated with synthesized peptideImmunohistochemical analysis of paraffin-embedded breast carcinoma, using ab47489 at a 1/50 dilution.
Left image: Untreated
Right image: Treated with synthesized peptide
ICC/IF image of ab47489 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab47489, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
ab47489 staining BCL2 in MCF7 cells treated with ICI 182,780 (ab120131), by ICC/IF. Decrease in BCL2 expression correlates with increased concentration of ICI 182,780 as described in literature. The cells were incubated at 37°C for 3h in media containing different concentrations of ab120131 (ICI 182,780) 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 ab47489 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A goat anti-rabbit DyLight 488 polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
ab47489 staining Bcl-2 MCF7 cells treated with diadzein (ab120391), by ICC/IF. Decrease in Bcl2 expression correlates with increased concentration of diadzein, as described in literature. The cells were incubated at 37°C for 6h in media containing different concentrations of ab120391 (diadzein) 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 ab47489 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A goat anti-rabbit DyLight 488 polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody.
References for Anti-Bcl-2 antibody (ab47489)
This product has been referenced in:
Loughran ST et al. Bfl-1 is a crucial pro-survival nuclear factor-?B target gene in Hodgkin/Reed-Sternberg cells. Int J Cancer129:2787-96 (2011).
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