Multiparameter flow cytometric analysis of CD5 expression on Human peripheral leucocyte populations. Human whole blood was stained with BD Horizon™ RY586 Mouse Anti-Human CD5 antibody (Cat. No. 568475/568476; Right Plot) or with BD Horizon™ RY586 Mouse IgG1, κ Isotype Control (Cat. No. 568097; Left Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing CD5 expression (or Ig Isotype control staining) versus side light scatter signals (SSC) was derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometry and data analysis were 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™ RY586 Mouse Anti-Human CD5 antibody (Cat. No. 568475/568476; Right Plot) or with BD Horizon™ RY586 Mouse IgG1, κ Isotype Control (Cat. No. 568097; Left Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing CD5 expression (or Ig Isotype control staining) versus side light scatter signals (SSC) was derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer and FlowJo™ software.
Product Details
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® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- CF™ is a trademark of Biotium, Inc.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
RY586 Mouse IgG1, κ Isotype Control RUO
Size
50 µg
Cat No.
568097
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
568476 Rev. 1
Antibody Details
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).
568476 Rev. 1
Format Details
RY586
The BD Horizon RealYellow™ 586 (RY586) Dye is part of the BD family of yellow-green dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 565-nm and an emission maximum (Em Max) at 586-nm. Driven by BD innovation, RY586 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY586 can be used as an alternative to PE and we recommend using an optical filter centered near 586-nm (eg, a 586/15-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE. Compared to PE, RY586 is similar in brightness, minimal spillover into Blue detectors, and increased spillover into the 610/20-nm (PE-CF594) detector. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.
RY586
Yellow-Green 561 nm
564 nm
586 nm
568476 Rev.1
Citations & References
Development References (8)
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Bernard A, Boumsell L, Dausset J, Milstein C, Schlossman SF, ed. Leukocyte Typing. New York: Springer-Verlag; 1984:1-814.
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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.
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In: Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007:49-50.
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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
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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
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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.
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McMichael AJ. A.J. McMichael .. et al., ed. Leucocyte typing III : white cell differentiation antigens. Oxford New York: Oxford University Press; 1987:1-1050.
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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
568476 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
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.
