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BD Pharmingen™ PE-Cy™5 Mouse Anti-Human CD21
Clone B-ly4 (RUO)
Flow cytometric analysis of CD21 expression on human peripheral blood lymphocytes. Whole blood was stained with either PE-Cy™5 Mouse Anti-Human CD21 (Cat. No. 551064; solid line histogram) or PE-Cy™5 Mouse IgG1 κ Isotype Control (Cat. No. 555750; dashed line histogram). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Fluorescent histograms were derived from gated events with the side and forward light-scattering characteristics of viable lymphocytes.
Flow cytometric analysis of CD21 expression on human peripheral blood lymphocytes. Whole blood was stained with either PE-Cy™5 Mouse Anti-Human CD21 (Cat. No. 551064; solid line histogram) or PE-Cy™5 Mouse IgG1 κ Isotype Control (Cat. No. 555750; dashed line histogram). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Fluorescent histograms were derived from gated events with the side and forward light-scattering characteristics of viable lymphocytes.
Product Details
BD Pharmingen™
CR2; Complement receptor type 2; C3DR; EBV-R; Epstein-Barr virus receptor
Human (QC Testing), Rhesus, Cynomolgus, Baboon, Pig (Tested in Development)
Mouse IgG1, κ
Flow cytometry (Routinely Tested)
20 µl
IV B98
AB_394028
Aqueous buffered solution containing BSA and ≤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 with PE-Cy5 (formerly known as BD Cy-Chrome™) under optimum conditions, and unconjugated antibody and free PE-Cy5 were removed.
Product Notices
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- PE-Cy5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the PE-Cy5 tandem fluorochrome, extra care must be taken when using dual-laser cytometers which may directly excite both PE and Cy5™.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- PE-Cy5 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by the 488 nm light of an Argon ion laser and serves as an energy donor, coupled to the cyanine dye Cy5, which acts as an energy acceptor and fluoresces at 670 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in PE-Cy5, thus maximizing its fluorescence emission intensity, minimizing residual emission from PE, and minimizing lot-to-lot variation.
- Cy is a trademark of GE Healthcare.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- 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.
- PE-Cy5 tandem fluorochromes have been reported to bind some classes of human macrophages and granulocytes via Fc receptors, and PE has been reported to bind to mouse B lymphocytes via Fc receptors. Preincubation of mouse leukocytes with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 can reduce the non-specific binding of PE-Cy5-conjugated reagents to mouse B cells. However, PE-Cy5 conjugated reagents should not be used to stain splenocytes of SJL, NOD, and MRL mice as B lymphocytes and/or other leukocytes have been reported to non-specifically stain regardless of the use of Mouse BD Fc Block™ (the CD72c complex has been implicated for PE-Cy5 binding in these strains). Reagents conjugated to PE, PerCP, PerCP-Cy5.5, APC, and APC-Cy7 tandem fluorochrome can be used on leukocytes from these mouse strains.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
551064 Rev. 8
Antibody Details
B-ly4
The B-ly4 monoclonal antibody specifically binds to CD21, a 145 kDa glycosylated type I integral membrane protein. CD21 is a receptor for the C3d complement fragment and for Epstein-Barr virus (EBV). CD21 is expressed on mature B cells, follicular dendritic cells, and some epithelial cells. It is also weakly expressed on the subset of mature T cells and thymocytes. CD21 plays a role in B-cell activation and proliferation. It may also play a role in modulating the function of T cells in the immune response to infections by lymphotropic viruses. Recently, CD21 was found to be part of a large complex containing CD19, CD81, and possibly other molecules.
This clone also cross-reacts with a major subset of, but not all, peripheral blood CD20+ lymphocytes of baboon, and both rhesus and cynomolgus macaque monkeys. A subset of CD3+ cells is also CD21+.
551064 Rev. 8
Format Details
PE-Cy5
PE-Cy5 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496 nm and 566 nm and an acceptor dye, Cy™5, with an emission maximum (Em Max) at 670-nm. PE designed to be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 670-nm (e.g., a 670/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and Yellow-Green (561-nm) lasers and the acceptor dye can be excited by the Red (627-640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE-Cy5
496 nm, 566 nm
670 nm
551064 Rev.8
Citations & References
View product citations for antibody "551064" on CiteAb
Development References (5)
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Fischer E, Delibrias C, Kazatchkine MD. Expression of CR2 (the C3dg/EBV receptor, CD21) on normal human peripheral blood T lymphocytes. J Immunol. 1991; 146(3):865-869. (Biology). View Reference
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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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Lin G-X, Yang X, Hollemweguer E, et al. Cross-reactivity of CD antibodies in eight animal species. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:519-523.
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Paterson RL, Kelleher C, Amankonah TD, et al. Model of Epstein-Barr virus infection of human thymocytes: expression of viral genome and impact on cellular receptor expression in the T-lymphoblastic cell line, HPB-ALL. Blood. 1995; 85(2):456-464. (Biology). View Reference
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Tsoukas CD, Lambris JD. Expression of EBV/C3d receptors on T cells: biological significance. Immunol Today. 1993; 14(2):56-59. (Biology). View Reference
551064 Rev. 8
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.
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.