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BV605 Mouse Anti-Human CD45RO
BV605 Mouse Anti-Human CD45RO
Multicolor flow cytometric analysis of CD45RO expression on human peripheral blood lymphocytes.  Left Panel: Whole blood was stained with BD Horizon™ BV605  Mouse Anti-Human CD45RO antibody (Cat. No. 562791/562790; solid line histogram), or with a BD Horizon™ BV605  Mouse IgG2a, κ Isotype Control (Cat. No. 562778; dashed line histogram). Right Panel: Whole blood was stained with BD Horizon™ BV605  Mouse Anti-Human CD45RO antibody in conjunction with a BD Horizon™ V450  Mouse Anti-Human CD45RA antibody (Cat. No. 560362/ 560363).      Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No.555899). The fluorescence histograms and dot plot (CD45RO versus CD45RA expression) were derived from 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.  Left Panel: Whole blood was stained with BD Horizon™ BV605  Mouse Anti-Human CD45RO antibody (Cat. No. 562791/562790; solid line histogram), or with a BD Horizon™ BV605  Mouse IgG2a, κ Isotype Control (Cat. No. 562778; dashed line histogram). Right Panel: Whole blood was stained with BD Horizon™ BV605  Mouse Anti-Human CD45RO antibody in conjunction with a BD Horizon™ V450  Mouse Anti-Human CD45RA antibody (Cat. No. 560362/ 560363).      Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No.555899). The fluorescence histograms and dot plot (CD45RO versus CD45RA expression) were derived from 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
AB_2744411
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 with BD Horizon™ BV605 under optimum conditions, and unconjugated antibody and free BD Horizon™ BV605 were removed.

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.

For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  4. 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.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. 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 BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
  8. CF™ is a trademark of Biotium, Inc.
562791 Rev. 3
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.

562791 Rev. 3
Format Details
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BV605
The BD Horizon Brilliant Violet™ 605 (BV605) dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 407-nm and an acceptor dye with an emission maximum (Em Max) at 605-nm. BV605, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 610-nm (e.g., a 610/20-nm bandpass filter). The acceptor dye can be excited by the yellow-green (561-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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BV605
Violet 405 nm
407 nm
605 nm
562791 Rev.3
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. (Immunogen: 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.
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562791 Rev. 3

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