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Does not react withRat
Synthetic peptide corresponding to Bcl-2 aa 41-54.
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.
|IHC-P||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|ICC/IF||Use a concentration of 5 µg/ml.|
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.
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