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. Other applications not tested.
Recommended Dilution:
Flow Cytometry: 1:50 (5ug/ml) labels 5x10e5 cells
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. 5 x 10e5 cells per tube were incubated 45 minutes on ice with 80ul of B0807-06G at a dilution of 1:50 (5ug/ml).
4. Cells were then washed three times, fixed and analyzed by FACS. Cells stained positive with a mean shift of 1.09 log10 fluorescent units when compared to a mouse IgG1/FITC negative control at a similar concentration.
5. Binding was blocked when cells were pre incubated 10 minutes with 20ul of 0.5mg/ml anti-bcl-2 antibody (Catalog # B0807-06E).
Storage and Stability:
May be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. FITC conjugates are sensitive to light.
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