Anti-Bcl-2 (B Cell Leukemia 2) (Biotin)

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 in Flow Cytometry, Western Blot and Immunohistochemistry. Other applications not tested. Recommended Dilutions: Flow Cytometry: 5ug/ml, 80ul labels 5x10e5 Nalm-6 tumor cells, Immunohistochemistry: Fixed, paraffin sections, Optimal dilutions to be determined by the researcher. Flow Cytometry Protocol: 1. Cultured Nalm-6 human tumor cells were fixed for 30 min 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. Five x 10e5 cells per tube were incubated 45 minutes on ice with 80ul of B0807-06F at 5ug/ml. , 4. Cells were then washed twice and incubated with S7973-88P Streptavidin (PE)., 5. Cells were washed three times, fixed and analyzed by FACS., 6. Cells stained with a mean shift of 0.39 log10 fluorescent units when compared to a Mouse IgG1 negative control (Biotin) at a similar concentration., 7. This shift was blocked when permeabalized cells were preincubated 10 minutes with 20ul of 0.5mg/ml B0807-06E. 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.