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RB670 Mouse Anti-Human CD45RA
RB670 Mouse Anti-Human CD45RA
Multicolor flow cytometric analysis of CD45RA expression on Human peripheral blood leukocytes.  Human whole blood was stained with BD Horizon™ RY703 Mouse Anti-Human CD45RO (Cat. No. 571463; Bottom Plots) and with either BD Horizon™ RB670 Mouse IgG1, κ Isotype Control (Cat. No. 571784; Left Plots) or BD Horizon™ RB670 Mouse Anti-Human CD45RA antibody (Cat. No. 571965/571966; Right Plots). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The bivariate pseudocolor density plots showing the correlated expression of CD45RA (or Ig Isotype control staining) versus either side light-scatter (SSC-A) signals (Top Plots) or CD45RO (Bottom Plots) were derived from gated events with the forward and side light-scatter characteristics of either viable leukocytes or lymphocytes, respectively. Samples were acquired using a BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.10 Software.
Multicolor flow cytometric analysis of CD45RA expression on Human peripheral blood leukocytes.  Human whole blood was stained with BD Horizon™ RY703 Mouse Anti-Human CD45RO (Cat. No. 571463; Bottom Plots) and with either BD Horizon™ RB670 Mouse IgG1, κ Isotype Control (Cat. No. 571784; Left Plots) or BD Horizon™ RB670 Mouse Anti-Human CD45RA antibody (Cat. No. 571965/571966; Right Plots). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The bivariate pseudocolor density plots showing the correlated expression of CD45RA (or Ig Isotype control staining) versus either side light-scatter (SSC-A) signals (Top Plots) or CD45RO (Bottom Plots) were derived from gated events with the forward and side light-scatter characteristics of either viable leukocytes or lymphocytes, respectively. Samples were acquired using a BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.10 Software.
Product Details
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BD Horizon™
CD45R; PTPRC; LCA; Leukocyte common antigen
Human (QC Testing), Rhesus,Cynomolgus,Baboon (Tested in Development)
Mouse IgG1, κ
Flow cytometry (Routinely Tested)
5 µl/test
5788
Aqueous buffered solution containing ≤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 unreacted dye was 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. 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.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
  9. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
  10. For U.S. patents that may apply, see bd.com/patents.
  11. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
  12. 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.
  13. Tandem fluorochromes contain both an energy donor and an energy acceptor. 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 the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
  14. Clones without a listed workshop have not been investigated in an HLDA workshop to receive a CD nomenclature. We use “CD” provisionally when our internal testing indicates that this clone binds to the same CD antigen as workshopped clones.
571966 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.

571966 Rev. 1
Format Details
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RB670
The BD Horizon RealBlue™ 670 (RB670) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 492 nm and an emission maximum (Em Max) at 670 nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB670 can be used on both spectral and conventional cytometers and is designed to be primarily excited by the Blue laser (488-nm). For conventional instruments equipped with only a Blue laser (488-nm), RB670 can be used as an alternative to PE-Cy5 and we recommend using an optical filter centered near 670-nm (eg, a 670/30-nm bandpass filter). For conventional and spectral instruments equipped with both a Blue (488-nm) and Yellow-Green (561-nm) laser and appropriate detectors, it can be used in conjunction with PE-Cy5.
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RB670
Blue 488 nm
492 nm
670 nm
571966 Rev.1
Citations & References
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View product citations for antibody "571966" on CiteAb

Development References (6)

  1. Arbore G, West EE, Rahman J, et al. Complement receptor CD46 co-stimulates optimal human CD8+ T cell effector function via fatty acid metabolism.. Nat Commun. 2018; 9(1):4186. (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. Palermo B, Franzese O, Donna CD, et al. Antigen-specificity and DTIC before peptide-vaccination differently shape immune-checkpoint expression pattern, anti-tumor functionality and TCR repertoire in melanoma patients.. Oncoimmunology. 2018; 7(12):e1465163. (Clone-specific: Flow cytometry). View Reference
  4. 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
  5. 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
  6. van der Windt DJ, Dons EM, Montoya CL, et al. T-lymphocyte homeostasis and function in infant baboons: implications for transplantation. Transplantation. 2012; 25(2):218-228. (Clone-specific: Flow cytometry). View Reference
View All (6) View Less
571966 Rev. 1

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