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R718 Mouse Anti-Human CD5
R718 Mouse Anti-Human CD5

Multiparameter flow cytometric analysis of CD5 expression on human peripheral leucocyte populations. Human whole blood was stained with BD Horizon™ R718 Mouse Anti-Human CD5 antibody (Cat. No. 567058/567237; Right Plot) or with a BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; Left Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Bivariate pseudocolor density plots showing CD5 expression (or Ig Isotype control staining) versus side light scatter signals (SSC) were derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer and FlowJo™ software.

Multiparameter flow cytometric analysis of CD5 expression on human peripheral leucocyte populations. Human whole blood was stained with BD Horizon™ R718 Mouse Anti-Human CD5 antibody (Cat. No. 567058/567237; Right Plot) or with a BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; Left Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Bivariate pseudocolor density plots showing CD5 expression (or Ig Isotype control staining) versus side light scatter signals (SSC) were derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer and FlowJo™ software.

Product Details
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BD Horizon™
CD5 antigen (p56-62); T1; Tp67; LEU1; Lymphocyte antigen T1/Leu-1
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human T cells
Flow cytometry (Routinely Tested)
5 µl
I T4; III T094, T518
921
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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  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. 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.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
567058 Rev. 1
Antibody Details
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UCHT2

The UCHT2 monoclonal antibody specifically binds to CD5. CD5 is a 67 kDa single-chain, type 1 transmembrane glycoprotein expressed on most thymocytes, the majority of peripheral T lymphocytes and a subpopulation of B cells. CD72 has been shown to be the natural ligand for CD5. CD5+ B cells produce polyreactive antibodies (mostly IgM).

The antibody was conjugated to BD Horizon Red 718, which has been developed exclusively by 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.

567058 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
567058 Rev.1
Citations & References
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Development References (8)

  1. Bernard A, Boumsell L, Dausset J, Milstein C, Schlossman SF, ed. Leukocyte Typing. New York: Springer-Verlag; 1984:1-814.
  2. Bernard A, Boumsell L, Hill C. Joint report of the first international workshop on human leucocyte differentiation antigens by the investigators of the participating laboratories: T2 protocol. In: Bernard A. A. Bernard .. et al., ed. Leucocyte typing : human leucocyte differentiation antigens detected by monoclonal antibodies : specification, classification, nomenclature = Typage leucocytaire : antigènes de différenciation leucocytaire humains révélés par les anticorps monoclonaux : "Rapports des études communes". Berlin New York: Springer-Verlag; 1984:25-60.
  3. In: Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007:49-50.
  4. Lankester AC, van Schijndel GM, Cordell JL, van Noesel CJ, van Lier RA. CD5 is associated with the human B cell antigen receptor complex. Eur J Immunol. 1994; 24(4):812-816. (Biology). View Reference
  5. Lydyard PM, Lamour A, MacKenzie LE, Jamin C, Mageed RA, Youinou P. CD5+ B cells and the immune system. Immunol Lett. 1993; 38(2):159-166. (Biology: Flow cytometry). View Reference
  6. McMichael AJ, Gotch FM. T-cell antigens: new and previously defined clusters. In: McMichael AJ. A.J. McMichael .. et al., ed. Leucocyte typing III : white cell differentiation antigens. Oxford New York: Oxford University Press; 1987:31-62.
  7. McMichael AJ. A.J. McMichael .. et al., ed. Leucocyte typing III : white cell differentiation antigens. Oxford New York: Oxford University Press; 1987:1-1050.
  8. Wallace DL, Beverley PC. Phenotypic changes associated with activation of CD45RA+ and CD45RO+ T cells. Immunology. 1990; 69(3):460-467. (Clone-specific: Flow cytometry). View Reference
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567058 Rev. 1

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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.