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PE-CF594 Mouse Anti-Human CD45RO
PE-CF594 Mouse Anti-Human CD45RO
Multicolor flow cytometric analysis of CD45RO expression on human peripheral blood lymphocytes. Human whole blood was stained with the BD Horizon™ PE-CF594 Mouse Anti-Human CD45RO antibody (Cat. No. 562299/562327; solid line histogram) and APC Mouse Anti-Human CD45RA antibody (Cat. No. 550855/561884). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The two color flow cytometric dot plots show the correlated expression of CD45RO versus CD45RA for gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Multicolor flow cytometric analysis of CD45RO expression on human peripheral blood lymphocytes. Human whole blood was stained with the BD Horizon™ PE-CF594 Mouse Anti-Human CD45RO antibody (Cat. No. 562299/562327; solid line histogram) and APC Mouse Anti-Human CD45RA antibody (Cat. No. 550855/561884). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The two color flow cytometric dot plots show the correlated expression of CD45RO versus CD45RA for gated 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 Horizon™
CD45R; PTPRC; LCA; Leukocyte common antigen; GP180; LY5; T200
Human (QC Testing)
Mouse BALB/c IgG2a, κ
Human IL-2-dependent T-cell line
Flow cytometry (Routinely Tested)
5 µl
IV N31
5788
AB_11154398
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 with BD Horizon™ PE-CF594 under optimum conditions, and unconjugated antibody and free PE-CF594 were 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. When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
  3. 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.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. CF™ is a trademark of Biotium, Inc.
  8. 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.
  9. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  10. 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.
  11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
562327 Rev. 1
Antibody Details
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UCHL1

The UCHL1 monoclonal antibody specifically binds to the 180 kDa isoform of CD45 (aka, the Leukocyte Common Antigen). CD45RO is a type I transmembrane glycoprotein that has cytoplasmic protein tyrosine phosphatase activity and functions in signal transduction pathways. This CD45 isoform does not include amino acid sequences encoded by the variable CD45 exons A, B, or C. CD45RO is expressed on most thymocytes, activated T cells, memory T cells, granulocytes and monocytes, but only on a proportion of resting T cells. CD45RO and CD45RA antibodies seem to define complementary, predominantly non-overlapping, populations in resting peripheral T cells, demonstrating heterogeneity within the CD8 and CD4 subpopulations. CD45RO binds to CD22.

This antibody is conjugated to BD Horizon™ PE-CF594, which has been developed exclusively by BD Biosciences as a better alternative to PE-Texas Red. PE-CF594 excites and emits at similar wavelengths to PE-Texas Red yet exhibits improved brightness and spectral characteristics. Due to PE having maximal absorption peaks at 496 nm and 564 nm, PE-CF594 can be excited by the blue (488-nm), green (532-nm) and yellow-green (561-nm) lasers and can be detected with the same filter set as PE-Texas Red (eg 610/20-nm filter).

562327 Rev. 1
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 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
562327 Rev.1
Citations & References
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Development References (5)

  1. Akbar AN, Terry L, Timms A, Beverley PC, Janossy G. Loss of CD45R and gain of UCHL1 reactivity is a feature of primed T cells. J Immunol. 1988; 140(7):2171-2178. (Clone-specific: Flow cytometry). View Reference
  2. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  3. Norton AJ, Ramsay AD, Smith SH, Beverley PC, Isaacson PG. Monoclonal antibody (UCHL1) that recognises normal and neoplastic T cells in routinely fixed tissues. J Clin Pathol. 1986; 39(4):399-405. (Clone-specific: Immunohistochemistry). View Reference
  4. Smith SH, Brown MH, Rowe D, Callard RE, Beverley PC. Functional subsets of human helper-inducer cells defined by a new monoclonal antibody, UCHL1. Immunology. 1986; 58(1):63-70. (Clone-specific: Flow cytometry, Immunohistochemistry, Immunoprecipitation). View Reference
  5. Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
View All (5) View Less
562327 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.