Description
Bcl-2 is considered to be novel among proto-oncogenes because it blocks apoptosis (programmed cell death) in many cell types. Apoptosis is an active form of cellular suicide that typically requires new RNA and protein synthesis and is associated with distinct morphological changes including cell shrinkage, cytoplasm membrane blebbing, nuclear fragmentation and DNA degradation. The Bcl-2 gene was first found in t(14:18) containing follicular B-cell lymphomas. A high proportion of these lymphomas contain t(14:18) chromosomal translocations involving the human Bcl-2 gene. Translocation of Bcl-2 sequences from chromosome 18 onto the transcriptionally active immunoglobulin locus at chromosome band 14q32 in B-cells deregulates Bcl-2 gene expression, resulting in high levels of Bcl-2 mRNA and protein expression. Because Bcl-2 blocks apoptosis it may contribute to tumorigenisis by prolonging cell survival rather than by accelerating the rate of cell proliferation. Mouse Bcl-2 migrates at a reduced molecular weight of ~26 kD. This antibody recognizes a 26 kD band representing the mouse p26-Bcl-2 protein. Additional minor bands at 27-31 kD and 18-21 kD may be visualized. The 27-31 kD upper band may represent a larger isoform, whereas the 18-21 kD lower band may be an internal translation or proteolytic product. 3F11 does not cross-react with human Bcl-2. For detection of human Bcl-2, refer to clone 6C8 (Cat. No. 551051), or clone 4D7 (Cat. No. 554202).
Reported ranges for usage in western blot and IHC on frozen sections, is around 5 µg/ml. The antibody should be titrated for each application. Thymus and M1 mouse myeloma cells (ATCC TIB-192) are suggested as positive controls. For immunofluroescent staining and flow cytometry, the directly conjugated formats of this clone are recommended (Cat No. 554221 and 556537).
Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
3. Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
