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Alexa Fluor® 647 Mouse Anti-Human Bcl-2
Alexa Fluor® 647 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 Alexa Fluor® 647 Mouse IgG1 κ Isotype control (Cat. No. 557732/557783; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human Bcl-2 antibody (Cat. No. 563600; 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 BD 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 Alexa Fluor® 647 Mouse IgG1 κ Isotype control (Cat. No. 557732/557783; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human Bcl-2 antibody (Cat. No. 563600; 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 BD LSRFortessa™ Cell Analyzer System.
Product Details
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BD Pharmingen™
BCL2; Apoptosis regulator Bcl-2; B-cell CLL/lymphoma 2
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human Bcl-2 synthetic peptide aa. 41-54
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_2738306
Aqueous buffered solution containing BSA and ≤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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Product Notices

  1. 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).
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  8. 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.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
563600 Rev. 1
Antibody Details
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Bcl-2/100

The Bcl-2/100 monoclonal antibody specifically binds to human Bcl-2. It has been reported not to crossreact with mouse Bcl-2. A synthetic peptide corresponding to amino acids 41-54 (GAAPAPGIFSSQPG) of human Bcl-2 was used as the immunogen. This peptide sequence reportedly is not conserved between human and mouse. Programmed cell death (apoptosis) is a normal physiologic process which occurs during embryonic development as well as in maintenance of tissue homeostasis. The apoptotic program is characterized by certain morphological features. These include changes in the plasma membrane such as loss of membrane asymmetry and attachment, a condensation of the cytoplasm and nucleus, and internucleosomal cleavage of DNA. In the final stages, the dying cells become fragmented into "apoptotic bodies" which are rapidly eliminated by phagocytic cells without eliciting significant inflammatory damage to surrounding cells. Members of the Bcl-2 family play a major role in regulating the response of cells to apoptotic signals. Bcl-2 is considered to be novel among proto-oncogenes because it blocks apoptosis in many cell types. Bcl-2 is thought to provide selective survival advantage for cells by blocking apoptosis and thus may contribute to tumorigenesis. Bcl-2 is an approximately 26 kDa intracellular, integral membrane protein found primarily in the nuclear envelope, endoplasmic reticulum and outer mitochondrial membrane.

563600 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
563600 Rev.1
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. (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
563600 Rev. 1

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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.