Multiparameter flow cytometric analysis of CD2 expression on Human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; Left Plot) or BD Horizon™ RB780 Mouse Anti-Human CD2 antibody (Cat. No. 569118/569119; Right Plot). Erythrocytes were lysed with BD FACS Lysing™ Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD2 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals 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 System and FlowJo™ software.
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Multiparameter flow cytometric analysis of CD2 expression on Human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon™ RB780 Mouse IgG1, κ Isotype Control (Cat. No. 568532; Left Plot) or BD Horizon™ RB780 Mouse Anti-Human CD2 antibody (Cat. No. 569118/569119; Right Plot). Erythrocytes were lysed with BD FACS Lysing™ Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD2 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals 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 System and FlowJo™ software.
Product Details
BD Horizon™
LFA-2; LFA-3 receptor; SRBC; T-cell surface antigen T11/Leu-5; erythrocyte receptor
Human (QC Testing), Rhesus,Cynomolgus,Baboon (Tested in Development)
Mouse BALB/c IgG1, κ
Human Phytohemagglutinin-treated Lymphoblasts
Flow cytometry (Routinely Tested)
5 µl/test
IV T085; VI 6T-078
914
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
- 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.
- 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.
- 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.
- 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.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
RB780 Mouse IgG1, κ Isotype Control RUO
Size
50 µg
Cat No.
568532
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Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
569119 Rev. 2
Antibody Details
RPA-2.10
The RPA-2.10 monoclonal antibody specifically binds to CD2 which is also known as Lymphocyte-function antigen-2 (LFA-2), LFA-3 receptor, Erythrocyte receptor, Sheep red blood cell (SRBC) receptor, or T-cell surface antigen T11/Leu-5. CD2 is a 50 kDa type I transmembrane glycoprotein. CD2 belongs to the immunoglobulin superfamily of proteins along with its primary ligand, LFA-3 (CD58). It is expressed on the surface of ~80-90% of human peripheral blood lymphocytes, greater than 95% of thymocytes, all T lymphocytes that form E-rosettes, and a subset of NK cells. CD2 functions as an adhesion receptor that binds to CD58 resulting in the activation of CD2-positive T cells and NK cells and in the regulation of their cytolytic activities.
569119 Rev. 2
Format Details
RB780
The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
RB780
Blue 488 nm
498 nm
781 nm
569119 Rev.2
Citations & References
View product citations for antibody "569119" on CiteAb
Development References (8)
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Aversa GG, Bishop GA, Suranyi MG, Hall BM. RPA-2.10: an anti-CD2 monoclonal antibody that inhibits alloimmune responses and monitors T cell activation. Transplant Proc. 1987; 19(1):277-278. (Immunogen: Flow cytometry, Immunoprecipitation, Inhibition). View Reference
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Bjornson-Hooper ZB, Fragiadakis GK, Spitzer MH, et al. A Comprehensive Atlas of Immunological Differences Between Humans, Mice, and Non-Human Primates.. Front Immunol. 2022; 13:867015. (Clone-specific: Flow cytometry). View Reference
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Hahn WC, Burakoff SJ, Bierer BE. Signal transduction pathways involved in T cell receptor-induced regulation of CD2 avidity for CD58. J Immunol. 1993; 150(7):2607-2619. (Biology). View Reference
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Jonker M, Slingerland W. Reactivity of mAb specific for human CD markers with Rhesus monkey leucocytes. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1058-1063.
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Kato K. CD2 Workshop Panel report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:39-43.
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Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
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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. (Clone-specific: Flow cytometry). View Reference
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Suranyi MG, Bishop GA, Clayberger C, et al. Lymphocyte adhesion molecules in T cell-mediated lysis of human kidney cells. Kidney Int. 1991; 39(2):312-319. (Clone-specific: Flow cytometry, Inhibition). View Reference
569119 Rev. 2
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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.