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568425_568426-1.png

Flow cytometric analysis of Bcl-2 expression in Mouse M1 myeloblast cells. Cells from the Mouse M1 (Myeloid leukemia, ATCC® TIB-192™) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) followed by washing and permeabilization using BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained in BD Perm/Wash™ Buffer with either Alexa Fluor™ 488 Hamster IgG, κ Isotype Control (Cat. No. 557674; dashed line histogram) or Alexa Fluor™ 488 Hamster Anti-Mouse Bcl-2 antibody (Cat. No. 568425/568426; solid line histogram) at 1 µg/test. Fluorescent histograms showing Bcl-2 expression (or Ig Isotype control staining) were derived from gated events with the side and forward light-scattering characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

568425_568426-1.png Green-Cap-Blue-Label.png

Flow cytometric analysis of Bcl-2 expression in Mouse M1 myeloblast cells. Cells from the Mouse M1 (Myeloid leukemia, ATCC® TIB-192™) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) followed by washing and permeabilization using BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained in BD Perm/Wash™ Buffer with either Alexa Fluor™ 488 Hamster IgG, κ Isotype Control (Cat. No. 557674; dashed line histogram) or Alexa Fluor™ 488 Hamster Anti-Mouse Bcl-2 antibody (Cat. No. 568425/568426; solid line histogram) at 1 µg/test. Fluorescent histograms showing Bcl-2 expression (or Ig Isotype control staining) were derived from gated events with the side and forward light-scattering characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Product Details
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BD Pharmingen™
B cell leukemia/lymphoma 2; Bcl-2; Bcl2; apoptosis regulator Bcl-2
Mouse (QC Testing), Human (Lack of Reactivity Confirmed in Development)
Armenian Hamster IgG1
Recombinant Mouse Bcl-2
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
12043
AB_3684261
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed.
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Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  8. For U.S. patents that may apply, see bd.com/patents.
  9. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  10. An isotype control should be used at the same concentration as the antibody of interest.
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568426 Rev. 1
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).

568426 Rev. 1
Format Details
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Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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Alexa Fluor™ 488
494 nm
517 nm
568426 Rev.1
Citations & References
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View product citations for antibody "568426" on CiteAb

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
568426 Rev. 1

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