Multicolor flow cytometric analysis of CD20 (cytoplasmic domain) expression in Human peripheral blood lymphocytes. Whole blood was treated with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) for 10 min at 37°C to lyse erythrocytes and fix the leucocytes in one step. The leucocytes were permeabilized with BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885) for 20 minutes. The cells were then stained with either BD Horizon™ R718 Mouse IgG2a, κ Isotype Control (Cat. No. 567089; dashed line histogram) or BD Horizon™ RY718 Mouse Anti-Human CD20 (cytoplasmic) antibody (Cat. No. 568050/568051; solid line histogram). The bivariate pseudocolor density plot showing the correlated expression of CD20 (cytoplasmatic domain) [or Ig Isotype control staining] vs side-light scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
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Multicolor flow cytometric analysis of CD20 (cytoplasmic domain) expression in Human peripheral blood lymphocytes. Whole blood was treated with BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) for 10 min at 37°C to lyse erythrocytes and fix the leucocytes in one step. The leucocytes were permeabilized with BD Phosflow™ Perm/Wash Buffer I (Cat. No. 557885) for 20 minutes. The cells were then stained with either BD Horizon™ R718 Mouse IgG2a, κ Isotype Control (Cat. No. 567089; dashed line histogram) or BD Horizon™ RY718 Mouse Anti-Human CD20 (cytoplasmic) antibody (Cat. No. 568050/568051; solid line histogram). The bivariate pseudocolor density plot showing the correlated expression of CD20 (cytoplasmatic domain) [or Ig Isotype control staining] vs side-light scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
Product Details
BD Horizon™
MS4A1; membrane-spanning 4-domains subfamily A member 1; B1; Bp35; LEU-16
Human (QC Testing)
Mouse BALB/c IgG2a, κ
Human B lymphoma cell line
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
V cB010
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.
- 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.
- 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.
- 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.
- This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
R718 Mouse IgG2a, κ Isotype Control RUO
Size
50 µg
Cat No.
567089
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Lyse/Fix Buffer 5X RUO
Size
250 mL
Cat No.
558049
Perm/Wash Buffer I RUO
Size
125 mL
Cat No.
557885
568051 Rev. 1
Antibody Details
H1
The H1 (FB1) antibody specificially binds to a cytoplasmic domain of CD20. CD20 is a 33-37-kDa four transmembrane phosphoprotein that is expressed by B lymphocytes from the pre-B stage and most malignant B cells and is lost during plasma cell differentiation. Low level CD20 expression is observed on a subset of normal circulating T lymphocytes, and CD20-positive T-cell lymphomas have been reported. The CD20 molecule is associated with membrane lipid raft domains, acts as a channel for calcium ions, and is involved in the regulation of B cell activation and survival. The cytoplasmic domain regions are serine and threonine rich and contain multiple phosphorylation consensus sequences.
568051 Rev. 1
Format Details
R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
R718
Red 627-640 nm
695 nm
718 nm
568051 Rev.1
Citations & References
View product citations for antibody "568051" on CiteAb
Development References (7)
-
Cragg MS, Walshe CA, Ivanov AO, Glennie MJ. The biology of CD20 and its potential as a target for mAb therapy. Curr Dir Autoimmun. 2005; 8:140-174. (Biology). View Reference
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Kitamura A, Yamashita Y, Mori N. CD20-positive cytotoxic T cell lymphoma: report of two cases and review of the literature. J Clin Exp Hematop. 2005; 45(1):45-50. (Biology).
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Nozawa Y, Abe M, Ohno H, Fukuhara S, Wakasa H. Production of two monoclonal antibodies (FB1 and FB21) useful for the identification of human B lymphocytes in formalin-fixed, paraffin-embedded tissues. J Pathol. 1994; 173:347-354. (Immunogen). View Reference
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Pylayeva-Gupta Y, Das S, Handler JS, et al. IL35-Producing B Cells Promote the Development of Pancreatic Neoplasia.. Cancer Discov. 2016; 6(3):247-55. (Clone-specific: Immunofluorescence, Immunohistochemistry). View Reference
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Sachen KL, Strohman MJ, Singletary J, et al. Self-antigen recognition by follicular lymphoma B-cell receptors.. Blood. 2012; 120(20):4182-90. (Clone-specific: Flow cytometry). View Reference
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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.
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Ziegler CGK, Kim J, Piersanti K, et al. Constitutive Activation of the B Cell Receptor Underlies Dysfunctional Signaling in Chronic Lymphocytic Leukemia.. Cell Rep. 2019; 28(4):923-937.e3. (Clone-specific: Flow cytometry). View Reference
568051 Rev. 1
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.