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R718 Mouse Anti-Human CD45RA
R718 Mouse Anti-Human CD45RA
Two-color flow cytometric analysis of CD45RA expression on human peripheral blood lymphocytes. Human whole blood was stained with FITC Mouse Anti-Human CD45RO antibody (Cat. No. 555492/561887), and with either BD Horizon™ R718 Mouse IgG2b, κ Isotype Control (Cat. No. 566952; Left Plot) or Mouse Anti-Human CD45RA antibody (Cat. No. 567073/567236; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). A bivariate pseudocolor density plot showing the correlated expression of CD45RA versus CD45RO was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
Two-color flow cytometric analysis of CD45RA expression on human peripheral blood lymphocytes. Human whole blood was stained with FITC Mouse Anti-Human CD45RO antibody (Cat. No. 555492/561887), and with either BD Horizon™ R718 Mouse IgG2b, κ Isotype Control (Cat. No. 566952; Left Plot) or Mouse Anti-Human CD45RA antibody (Cat. No. 567073/567236; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). A bivariate pseudocolor density plot showing the correlated expression of CD45RA versus CD45RO was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
Product Details
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BD Horizon™
CD45R; PTPRC; LCA; Leukocyte common antigen
Human (QC Testing)
Mouse IgG2b, κ
Flow cytometry (Routinely Tested)
5 µl
IV N906
5788
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 to the dye under optimum conditions and unreacted dye was 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 CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.

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. 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.
  4. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  5. This product is provided under an Agreement between BIOTIUM and BD Biosciences. 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.
  6. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
567236 Rev. 1
Antibody Details
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HI100

The HI100 monoclonal antibody specifically binds to the 220 kDa isoform of the human leukocyte common antigen, CD45RA. CD45RA is expressed on approximately 40-50% of peripheral CD4+ T cells, 50% of peripheral CD8+ T cells and on a portion of B cells and monocytes. The CD45RA antigen is expressed by naïve and activated T cells. CD45RA-specific antibodies are useful for the study of the suppressor/inducer subpopulation of CD4+ lymphocytes.

The antibody was conjugated to BD Horizon Red 718, which has been developed exclusively for BD Biosciences as a better alternative to Alexa Fluor™ 700. BD Horizon Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor™ 700.

567236 Rev. 1
Format Details
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R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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R718
Red 627-640 nm
695 nm
718 nm
567236 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. Koristka S, Cartellieri M, Theil A, et al. Retargeting of human regulatory T cells by single-chain bispecific antibodies.. J Immunol. 2012; 188(3):1551-8. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
  3. Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
567236 Rev. 1

Please refer to Support Documents for Quality Certificates


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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.