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PE-CF594 Mouse Anti-Human Bcl-2
PE-CF594 Mouse Anti-Human Bcl-2
Flow cytometric analysis of Bcl-2 expression in human peripheral blood mononuclear cells. Human PBMC were fixed and permeabilized using BD Cytofix/Cytoperm™ (Cat. No. 554714) followed by staining with either BD Horizon™ PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292; dashed line histogram) or BD Horizon™ PE-CF594 Mouse Anti-Human Bcl-2 antibody (Cat. No. 563601; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a LSRFortessa™ Cell Analyzer System.
Flow cytometric analysis of Bcl-2 expression in human peripheral blood mononuclear cells. Human PBMC were fixed and permeabilized using BD Cytofix/Cytoperm™ (Cat. No. 554714) followed by staining with either BD Horizon™ PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292; dashed line histogram) or BD Horizon™ PE-CF594 Mouse Anti-Human Bcl-2 antibody (Cat. No. 563601; solid line histogram). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a LSRFortessa™ Cell Analyzer System.
Product Details
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BD Horizon™
BCL2; Apoptosis regulator Bcl-2; B-cell CLL/lymphoma 2
Human (QC Testing)
Mouse IgG1, κ
Human Bcl-2 synthetic peptide aa. 41-54
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
596
AB_2738307
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

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 unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

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.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  9. CF™ is a trademark of Biotium, Inc.
  10. When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
  11. This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  12. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
  13. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
  14. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  15. For U.S. patents that may apply, see bd.com/patents.
563601 Rev. 3
Antibody Details
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Bcl-2/100

The Bcl-2/100 monoclonal antibody specifically recognizes human Bcl-2, also known as B-cell CLL/lymphoma 2. A synthetic peptide corresponding to amino acids 41-54 (GAAPAPGIFSSQPG) of human Bcl-2 was used as the immunogen. This ~26 kDa intracellular protein is encoded by BCL2 (BCL2 apoptosis regulator) which belongs to the Bcl-2 family of regulatory proteins. Bcl-2 is expressed in the endoplasmic reticulum, the outer mitochondrial membrane, and the nuclear envelope. Bcl-2 inhibits apoptosis and is involved in autophagy signaling. It is variably expressed in different cell types including hematopoietic progenitor cells, cells belonging to the T cell and B cells lineages, neurons, epithelial cells, endothelial cells, and melanocyte progenitors. Bcl-2 can form homodimers or heterodimers with other Bcl-2 family members that either block or promote apoptosis. Abnormal expression of Bcl-2 is associated with certain cancers including prostate cancer and B-cell lymphoma.

563601 Rev. 3
Format Details
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PE-CF594
BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, is designed to be excited by the blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the green (532-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE-CF594
Yellow-Green 561 nm
496 nm, 566 nm
615 nm
563601 Rev.3
Citations & References
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View product citations for antibody "563601" on CiteAb

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. (Immunogen: 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
563601 Rev. 3

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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.