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BB700 Mouse Anti-Human CD21
BB700 Mouse Anti-Human CD21
Two-color flow cytometric analysis of CD21 expression on human peripheral blood lymphocytes. Whole human blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD19 antibody (Cat. No. 562440/562441) and either BD Horizon™ BB700 Mouse IgG1, κ Isotype Control (Cat. No. 566404; Left Plot) or BD Horizon BB700 Mouse Anti-Human CD21 antibody (Cat. No. 566569/566570; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The two-color flow cytometric contour plot showing the correlated expression of CD21 (or Ig Isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD FACSCelesta™ Flow Cytometer System.
Two-color flow cytometric analysis of CD21 expression on human peripheral blood lymphocytes. Whole human blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD19 antibody (Cat. No. 562440/562441) and either BD Horizon™ BB700 Mouse IgG1, κ Isotype Control (Cat. No. 566404; Left Plot) or BD Horizon BB700 Mouse Anti-Human CD21 antibody (Cat. No. 566569/566570; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). The two-color flow cytometric contour plot showing the correlated expression of CD21 (or Ig Isotype control staining) versus CD19 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD FACSCelesta™ Flow Cytometer System.
Product Details
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BD Horizon™
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)
5 µl
IV B98
1380
AB_2739743
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 with BD Horizon™ BB700 under optimum conditions, and unconjugated antibody and free BD Horizon BB700 were removed.

Recommended Assay Procedures

     For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet for the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

     When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.

     For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  6. BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
  7. Cy is a trademark of GE Healthcare.
  8. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Antibody Details
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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.

The antibody was conjugated to BD Horizon BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes.   It is a polymer-based tandem dye developed exclusively by BD Biosciences.  With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.

Format Details
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BB700
The BD Horizon Brilliant™ Blue 700 (BB700) Dye is part of the BD Horizon Brilliant™ Blue family of dyes. This tandem fluorochrome is comprised of a polymer-technology dye donor with an excitation maximum (Ex Max) of 476-nm and an acceptor dye with an emission maximum (Em Max) at 695-nm. Driven by BD innovation, BB700 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 695-nm (e.g., a 695/20-nm bandpass filter). The donor dye can be excited by the Violet (405 nm) laser and the acceptor dye can be excited by the red (627–640 nm) laser resulting in cross-laser excitation and fluorescence spillover. BB700 Reagents are significantly brighter than equivalent PerCP or PerCP-Cy5.5 reagents and are less sensitive to photobleaching. In addition, BB700 shows much less excitation by the violet (407-nm) laser resulting in less spillover. BB700 has minimal yellow green (562-nm) excitation and is ideal for instruments with both blue (488-nm) and yellow green (562-nm) lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BB700
Blue 488 nm
476 nm
695 nm
Citations & References
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Development References (7)

  1. Fearon DT. The CD19-CR2-TAPA-1 complex, CD45 and signaling by the antigen receptor of B lymphocytes. Curr Opin Immunol. 1993; 5(3):341-348. (Biology). View Reference
  2. 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
  3. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  4. 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.
  5. 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
  6. Timens W, Boes A, Vos H, Poppema S. Tissue distribution of the C3d/EBV-receptor: CD21 monoclonal antibodies reactive with a variety of epithelial cells, medullary thymocytes, and peripheral T-cells. Histochemistry. 1991; 95(6):605-611. (Clone-specific: Immunocytochemistry (cytospins), Immunohistochemistry). View Reference
  7. Tsoukas CD, Lambris JD. Expression of EBV/C3d receptors on T cells: biological significance. Immunol Today. 1993; 14(2):56-59. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.