Technical Data
B0807-06E
Bcl-2 (B Cell Leukemia 2)
Description:
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. Five 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 2 reagent IgG (FITC) PAB Gt xMo 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 4C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile 40-50% glycerol, aliquot and store at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
MabIgG17H161Affinity Purified
SizeStorageShippingSourceHost
100ug4C (-20C Glycerol)Blue IceHumanMouse
Concentration:
~1mg/ml
Immunogen:
Human bcl-2 peptide 41-54
Purity:
Purified by Protein A affinity chromatography.
Form
Supplied as a liquid in PBS, 0.5mg/ml gentamicin sulfate.
Specificity:
Recognizes the intracellular human bcl-2 protein at ~26kD.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
1. F. Pezzella, et al, (1990) Am J Pathol 137: 225-232. 2. I.A. Doussis, et al, (1993) Am J Clin Pathol 99: 663-667. 3. S. Cory, (1995) Annu Rev Immunol 13: 513-543.