Anti-Bcl-2 (B Cell Leukemia 2)

The human bcl-2 gene family plays a central role in lymphoid system cell survival. Continuous expression of bcl-2 protein is critical for preventing apoptosis of mature peripheral B and T cells. Applications: Suitable for use Flow Cytometry, Western Blot and Immunohistochemistry. Other applications not tested. Recommended Dilution: Flow Cytometry: 5mg/ml 80ul labels 5x10e5 cells, Optimal dilutions to be determined by the researcher. FACS Protocol: 1. Cultured Nalm-6 human tumor cells were fixed for 30 minutes with 2% buffered formaldehyde after which they were washed two times. , 2. Subsequent reagent incubations and washes were done using a buffer containing 0.3% Saponin to permeabolize cells. , 3. 5 x 10e5 cells per tube were preincubated ~5 minutes with 20ul of 250ug/ml human IgG (to block non specific binding). , 4. They were then incubated for 45 minutes on ice with 80ul of B0807-06E at 5ug/ml. , 5. Cells were then washed twice and incubated with FITC-conjugated polyclonal goat anti-mouse IgG secondary antibody, then washed three times, fixed and analyzed using a BD FACS Calibur. Cells stained with a mean shift of 0.56 log10 fluorescent units when compared to a Mouse IgG1 negative control at a similar concentration. Recommended Secondary Antibodies: I1904-06F: IgG, H&L (FITC) (X-Adsorbed) Pab Gt xMo, I1905-18D IgM (X-Adsorbed) (FITC) Pab GtxMo, Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.