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Purified Mouse Anti-Human Bcl-2 w/Control

Purified Mouse Anti-Human Bcl-2 w/Control

(RUO)
Purified Mouse Anti-Human Bcl-2 w/Control
Profile of permeabilized lymphocytes analyzed on a FACScan (BDIS, San Jose, CA) (left panel). Cells were stained with anti-human Bcl-2 (clone Bcl-2/100) or a mouse IgG1 isotype (negative) control followed by FITC-conjugated second step.
Purified Mouse Anti-Human Bcl-2 w/Control

Western blot analysis of Bcl-2 (right panel). Lysate from Jurkat cells was probed with clone Bcl-2/100 at concentrations of 1.0 (lane 1), 0.5 (lane 2), and 0.125 µg/ml (lane 3). Bcl-2 is identified as a band of ~26 kDa.

Purified Mouse Anti-Human Bcl-2 w/Control

Formalin-fixed, paraffin-embedded tissue sections stained for Bcl-2 expression using clone Bcl-2/100 and the SA-HRP method. (Left panel) Normal tonsil. (Right panel) Follicular lymphoma. In normal tonsil the mantle zone and intrafollicular region are stained; only occasional Bcl-2 positive cells are seen in the germinal center. In follicular lymphoma, tumor follicles are Bcl-2 positive.

Profile of permeabilized lymphocytes analyzed on a FACScan (BDIS, San Jose, CA) (left panel). Cells were stained with anti-human Bcl-2 (clone Bcl-2/100) or a mouse IgG1 isotype (negative) control followed by FITC-conjugated second step.

Western blot analysis of Bcl-2 (right panel). Lysate from Jurkat cells was probed with clone Bcl-2/100 at concentrations of 1.0 (lane 1), 0.5 (lane 2), and 0.125 µg/ml (lane 3). Bcl-2 is identified as a band of ~26 kDa.

Formalin-fixed, paraffin-embedded tissue sections stained for Bcl-2 expression using clone Bcl-2/100 and the SA-HRP method. (Left panel) Normal tonsil. (Right panel) Follicular lymphoma. In normal tonsil the mantle zone and intrafollicular region are stained; only occasional Bcl-2 positive cells are seen in the germinal center. In follicular lymphoma, tumor follicles are Bcl-2 positive.

Product Details
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BD Pharmingen™
Human (QC Testing)
Western blot (Routinely Tested), Flow cytometry, Immunohistochemistry-formalin (antigen retrieval required), Immunohistochemistry-frozen (Tested During Development)
RUO


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. Because Bcl-2 blocks apoptosis it may contribute to tumorigenisis by prolonging cell survival, rather than by accelerating the rate of cell proliferation. Human Bcl-2 protein migrates at a molecular weight of ~26 kDa by SDS-PAGE. Bcl-2/100 recognizes a 26 kDa band representing human Bcl-2. Additional minor bands at 27-31 kDa and 18-21 kDa may also be visualized. The 27-31 kDa upper band may represent a larger isoform, whereas the 18-21 kDa lower band may be an internal translation or proteolytic product, therefore, a synthetic peptide corresponding to amino acids 41-54 (GAAPAPGIFSSQPG) of human Bcl-2 was used as immunogen. This peptide sequence is not conserved between human and mouse. Bcl-2/100 does not cross-react with mouse Bcl-2. For detection of mouse Bcl-2 refer to clone 3F11 (Cat. No. 554218), polyclonal rabbit anti-rat/mouse Bcl-2 antiserum (Cat. No. 554087), and polyclonal rabbit anti-mouse Bcl-2 antiserum (Cat. No. 554279).

Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at -20°C.

Recommended Assay Procedures

Western Blot: Jurkat control lysate [50 µg (1 µg/µl)] is provided as a western blot positive control (Comp. No. 51-16526N; Store lysate at -20°C). Additional control lysate (Cat. No. 611451) is sold separately.

IHC: For paraffin-embedded sections, tissues should be fixed in either Bouin's fixative or formalin.

For immunofluorescent staining and flow cytometry,  our directly conjugated formats are recommended, Cat. No. 556535 and 556357.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
551107 Rev. 5
Components
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Description Quantity/Size Part Number EntrezGene ID
Purified Mouse Anti-Human Bcl-2 50 µg (3 ea) 51-6511GR N/A
Jurkat Cell Lysate 50 µg (1 ea) 51-16526N N/A
551107 Rev. 5
Citations & References
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Development References (7)

  1. 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
  2. Pezzella F, Jones M, Ralfkiaer E, Ersbøll J, Gatter KC, Mason DY. Evaluation of bcl-2 protein expression and 14;18 translocation as prognostic markers in follicular lymphoma. Br J Cancer. 1992; 65(1):87-89. (Clone-specific: Immunohistochemistry). View Reference
  3. Pezzella F, Tse AG, Cordell JL, Pulford KA, Gatter KC, Mason DY. Expression of the bcl-2 oncogene protein is not specific for the 14;18 chromosomal translocation. Am J Pathol. 1990; 137(2):225-232. (Clone-specific: Immunohistochemistry, Western blot). View Reference
  4. Pezzella F, Turley H, Kuzu I, et al. bcl-2 protein in non-small-cell lung carcinoma. N Engl J Med. 1993; 329(10):690-694. (Clone-specific: Immunohistochemistry). View Reference
  5. Reed JC, Meister L, Tanaka S, et al. Differential expression of bcl2 protooncogene in neuroblastoma and other human tumor cell lines of neural origin. Cancer Res. 1991; 51(24):6529-6538. (Biology). View Reference
  6. Tanaka S, Saito K, Reed JC. Structure-function analysis of the Bcl-2 oncoprotein. Addition of a heterologous transmembrane domain to portions of the Bcl-2 beta protein restores function as a regulator of cell survival. J Biol Chem. 1993; 268(15):10920-10926. (Biology). View Reference
  7. Williams GT. Programmed cell death: apoptosis and oncogenesis. Cell. 1991; 65(7):1097-1098. (Biology). View Reference
View All (7) View Less
551107 Rev. 5

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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.