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BD Horizon™ BB515 Mouse Anti-Human CD20
Clone 2H7 (RUO)
Two-color flow cytometric analysis of CD20 expression on human peripheral blood lymphocytes - Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies. Human whole blood was stained with APC Mouse Anti-Human CD19 antibody (Cat. No. 555415/561742) and either BD Horizon BB515 Mouse IgG2b, κ Isotype Control (Cat. No. 564510; Left Panel), BD Horizon BB515 Mouse Anti-Human CD20 antibody (Cat. No. 564568/564569; Middle Panel), or FITC Mouse Anti-Human CD20 antibody (Cat. No. 555622/556632/560962; Right Panel). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No.349202). Two-color flow cytometric contour plots showing the correlated expression of CD20 (or Ig Isotype control staining) versus CD19 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of CD20 expression on human peripheral blood lymphocytes - Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies. Human whole blood was stained with APC Mouse Anti-Human CD19 antibody (Cat. No. 555415/561742) and either BD Horizon BB515 Mouse IgG2b, κ Isotype Control (Cat. No. 564510; Left Panel), BD Horizon BB515 Mouse Anti-Human CD20 antibody (Cat. No. 564568/564569; Middle Panel), or FITC Mouse Anti-Human CD20 antibody (Cat. No. 555622/556632/560962; Right Panel). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No.349202). Two-color flow cytometric contour plots showing the correlated expression of CD20 (or Ig Isotype control staining) versus CD19 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of CD20 expression on human peripheral blood lymphocytes - Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies. Human whole blood was stained with APC Mouse Anti-Human CD19 antibody (Cat. No. 555415/561742) and either BD Horizon BB515 Mouse IgG2b, κ Isotype Control (Cat. No. 564510; Left Panel), BD Horizon BB515 Mouse Anti-Human CD20 antibody (Cat. No. 564568/564569; Middle Panel), or FITC Mouse Anti-Human CD20 antibody (Cat. No. 555622/556632/560962; Right Panel). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No.349202). Two-color flow cytometric contour plots showing the correlated expression of CD20 (or Ig Isotype control staining) versus CD19 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of CD20 expression on human peripheral blood lymphocytes - Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies. Human whole blood was stained with APC Mouse Anti-Human CD19 antibody (Cat. No. 555415/561742) and either BD Horizon BB515 Mouse IgG2b, κ Isotype Control (Cat. No. 564510; Left Panel), BD Horizon BB515 Mouse Anti-Human CD20 antibody (Cat. No. 564568/564569; Middle Panel), or FITC Mouse Anti-Human CD20 antibody (Cat. No. 555622/556632/560962; Right Panel). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No.349202). Two-color flow cytometric contour plots showing the correlated expression of CD20 (or Ig Isotype control staining) versus CD19 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
BD Horizon™
MS4A1; B1; Bp35; LEU-16; S7
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse C57BL/6 IgG2b, κ
Human 6.16c1.3 B cell line
Flow cytometry (Routinely Tested)
5 µl
II B22; III B739, NL382; IV B201
AB_2744324
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™ BB515 under optimum conditions and unconjugated antibody was removed.
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
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 of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence 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
- 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.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 ML
Cat No.
554657
Brilliant Stain Buffer RUO
Size
100 Tests
Cat No.
563794
Lysing Solution 10X Concentrate CE_IVD
Size
100 ML
Cat No.
349202
Lysing Buffer RUO
Size
100 ML
Cat No.
555899
BB515 Mouse IgG2b, κ Isotype Control RUO
Size
50 µg
Cat No.
564510
564568 Rev. 3
Antibody Details
2H7
The 2H7 monoclonal antibody specifically binds to CD20, encoded by the MS4A1 (Membrane-spanning 4-domains, subfamily A, member 1) gene. CD20 is a 33-37 kDa, unglycosylated four-transmembrane phosphoprotein. CD20 is expressed on pre-B-cells, resting and activated B cells, and follicular dendritic cells, but not plasma cells. Low level CD20 expression is observed on a small subset of normal circulating T lymphocytes. The CD20 molecule is involved in the regulation of B-cell activation.
The antibody was conjugated to BD Horizon BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.
564568 Rev. 3
Format Details
BB515
The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye. It is not recommended to use BB515 in cocktails that include streptavidin conjugates as it may cause high background.
BB515
490 nm
515 nm
564568 Rev.3
Citations & References
View product citations for antibody "564568" on CiteAb
Development References (7)
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Clark EA, Yokochi T. Human B cell and B cell blast-associated surface molecules defined with monoclonal antibodies. In: Bernard A, Boumsell L, Dausset J, Milstein C, Schlossman SF, ed. Leukocyte Typing. Berlin: Springer-Verlag; 1984:339-346.
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Hultin LE, Hausner MA, Hultin PM, Giorgi JV. CD20 (pan-B cell) antigen is expressed at a low level on a subpopulation of human T lymphocytes. Cytometry. 1993; 14(2):193-204. (Clone-specific: Flow cytometry). 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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Ledbetter JA, Clark EA. Surface phenotype and function of tonsillar germinal center and mantle zone B cell subsets. Hum Immunol. 1986; 15:30-43. (Immunogen: Blocking, Flow cytometry). View Reference
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Loken MR, Shah VO, Dattilio KL, Civin CI. Flow cytometric analysis of human bone marrow. II. Normal B lymphocyte development. Blood. 1987; 70(5):1316-1324. (Biology). View Reference
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Reinherz EL. Ellis L. Reinherz .. et al., ed. Leukocyte typing II. New York: Springer-Verlag; 1986:1-560.
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Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
564568 Rev. 3
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.