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PE-Cy™7 Mouse Anti-Human CD45RA
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PE-Cy™7 Mouse Anti-Human CD45RA
Flow cytometric analysis of CD45RA expression on Rhesus macaque peripheral blood lymphocytes. Rhesus macaque whole blood was stained with PE-Cy™7 Mouse Anti-Human CD45RA antibody (Cat. No. 561216; solid line histogram) or with a PE-Cy™7 Mouse IgG1, κ Isotype Control (Cat. No. 557872; dashed line histogram). The erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of CD45RA expression on Rhesus macaque peripheral blood lymphocytes. Rhesus macaque whole blood was stained with PE-Cy™7 Mouse Anti-Human CD45RA antibody (Cat. No. 561216; solid line histogram) or with a PE-Cy™7 Mouse IgG1, κ Isotype Control (Cat. No. 557872; dashed line histogram). The erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Pharmingen™
Rhesus, Cynomolgus, Baboon (QC Testing)
Mouse IgG1, κ
Flow cytometry (Routinely Tested)
5 µl
5788
AB_10611721
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 with PE-Cy7 under optimum conditions, and unconjugated antibody and free PE-Cy7 were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

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. An isotype control should be used at the same concentration as the antibody of interest.
  3. Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
  4. Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
  5. This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
  6. PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  7. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. 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.
  10. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
561216 Rev. 1
Antibody Details
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5H9

The 5H9 monoclonal antibody specifically binds to the human form of the leukocyte common antigen (LCA), CD45RA. This clone also crossreacts with CD45RA expressed by peripheral blood lymphocytes, monocytes and some granulocytes from rhesus macaque, baboon, and cynomolgus monkeys. The observed frequency of CD45RA positive lymphocytes is higher in the non-human primate compared to what is seen with normal human donor lymphocytes.

561216 Rev. 1
Format Details
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PE-Cy7
PE-Cy7 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, Cy™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-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 Red (627–640-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-Cy7
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
781 nm
561216 Rev.1
Citations & References
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Development References (3)

  1. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  2. Reimann KA, Waite BC, Lee-Parritz DE, et al. Use of human leukocyte-specific monoclonal antibodies for clinically immunophenotyping lymphocytes of rhesus monkeys. Cytometry. 1994; 17(1):102-108. (Biology). View Reference
  3. Sopper S, Stahl-Hennig C, Demuth M, Johnston IC, Dorries R, ter Meulen V. Lymphocyte subsets and expression of differentiation markers in blood and lymphoid organs of rhesus monkeys. Cytometry. 1997; 29(4):351-362. (Biology). View Reference
561216 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.