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Purified Hamster Anti-Mouse Bcl-2
Purified Hamster Anti-Mouse Bcl-2
Western blot analysis of Bcl-2 expression on human and mouse thymocytes. Total organ lysates were probed with Purified Hamster Anti-Mouse Bcl-2 (Clone 3F11, Cat. No. 554218) or Purified Hamster Anti-Human Bcl-2 (Clone 6C8, Cat. No. 551051) and visualized with Horseradish Peroxidase (HRP) Mouse Anti-Armenian and Syrian Hamster IgG Cocktail (Cat. No. 554012). Clone 3F11 recognizes mouse (lane 1) but not human (lane 2) Bcl-2, while clone 6C8 recognizes human (lane 4) but not mouse Bcl-2 (lane 3).
Western blot analysis of Bcl-2 expression on human and mouse thymocytes. Total organ lysates were probed with Purified Hamster Anti-Mouse Bcl-2 (Clone 3F11, Cat. No. 554218) or Purified Hamster Anti-Human Bcl-2 (Clone 6C8, Cat. No. 551051) and visualized with Horseradish Peroxidase (HRP) Mouse Anti-Armenian and Syrian Hamster IgG Cocktail (Cat. No. 554012). Clone 3F11 recognizes mouse (lane 1) but not human (lane 2) Bcl-2, while clone 6C8 recognizes human (lane 4) but not mouse Bcl-2 (lane 3).
Product Details
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BD Pharmingen™
Mouse (QC Testing)
Armenian Hamster IgG1
Recombinant Mouse Bcl-2
Western blot (Routinely Tested), Fluorescence microscopy, Immunohistochemistry-frozen, Immunoprecipitation, Intracellular staining (flow cytometry) (Reported)
26 kDa
0.5 mg/ml
AB_395311
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

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).

Product Notices

  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.
554218 Rev. 7
Antibody Details
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3F11

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).

554218 Rev. 7
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
554218 Rev.7
Citations & References
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Development References (10)

  1. Hockenbery D, Nuñez G, Milliman C, Schreiber RD, Korsmeyer SJ. Bcl-2 is an inner mitochondrial membrane protein that blocks programmed cell death. Nature. 1990; 348(6299):334-336. (Biology). View Reference
  2. Krajewski S, Tanaka S, Takayama S, Schibler MJ, Fenton W, Reed JC. Investigation of the subcellular distribution of the bcl-2 oncoprotein: residence in the nuclear envelope, endoplasmic reticulum, and outer mitochondrial membranes. Cancer Res. 1993; 53(19):4701-4714. (Biology). View Reference
  3. Miyashita T, Krajewski S, Krajewska M, et al. Tumor suppressor p53 is a regulator of bcl-2 and bax gene expression in vitro and in vivo. Oncogene. 1994; 9(6):1799-1805. (Biology). View Reference
  4. Novack DV, Korsmeyer SJ. Bcl-2 protein expression during murine development. Am J Pathol. 1994; 145(1):61-73. (Clone-specific: Immunohistochemistry, Western blot). View Reference
  5. Oltvai ZN, Milliman CL, Korsmeyer SJ. Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell. 1993; 74(4):609-619. (Clone-specific: Immunoprecipitation). View Reference
  6. Reed JC, Tsujimoto Y, Alpers JD, Croce CM, Nowell PC. Regulation of bcl-2 proto-oncogene expression during normal human lymphocyte proliferation. Science. 1987; 236(4806):1295-1299. (Biology). View Reference
  7. Tsujimoto Y, Cossman J, Jaffe E, Croce CM. Involvement of the bcl-2 gene in human follicular lymphoma. Science. 1985; 228(4706):1440-1443. (Biology). View Reference
  8. Veis DJ, Sentman CL, Bach EA, Korsmeyer SJ. Expression of the Bcl-2 protein in murine and human thymocytes and in peripheral T lymphocytes. J Immunol. 1993; 151(5):2546-2554. (Immunogen: Flow cytometry, Fluorescence microscopy, Immunohistochemistry, Western blot). View Reference
  9. Veis DJ, Sorenson CM, Shutter JR, Korsmeyer SJ. Bcl-2-deficient mice demonstrate fulminant lymphoid apoptosis, polycystic kidneys, and hypopigmented hair. Cell. 1993; 75(2):229-240. (Clone-specific: Western blot). View Reference
  10. Williams GT. Programmed cell death: apoptosis and oncogenesis. Cell. 1991; 65(7):1097-1098. (Biology). View Reference
View All (10) View Less
554218 Rev. 7

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

Comparisons, where applicable, are made against older BD technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.