Preparation And Storage
Recommended Assay Procedures
Bead-based compensation or unmixing controls, such as BD® CompBeads or BD™ SpectraComp™, can be used as surrogates to assess fluorescence spillover when bound to fluorochrome-conjugated antibodies. Although these beads have spectral properties similar to cells, variations in spectral emission may occur, resulting in differing spillover values compared to biological controls. Therefore, it is considered best practice to compare the spillover obtained from cells and bead-based controls when using BD® CompBeads or BD™ SpectraComp™ for the first time, to ensure they are appropriate for the intended application.
For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. 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).
Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- 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.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
The GA-R2 (also known as HIR2) monoclonal antibody specifically binds to CD235a and CD235b. CD235a is also known as Glycophorin A (GYPA, GPA, GLPA), Sialoglycoprotein alpha, MN sialoglycoprotein, or PAS-2. CD235b is otherwise known as Glycophorin B (GYPB, GPB, GLPB), Sialoglycoprotein delta, SS-active sialoglycoprotein, or PAS-3. CD235a and CD235b are type I transmembrane sialoglycoproteins that are expressed on human erythrocytes, erythroid precursor cells and certain leukemic cell types. CD235a carries blood group M and N antigens, whereas CD235b contains S, s, and U antigens. This antibody is useful for the identification and characterization of erythrocytes, certain myeloid leukemic cell types, and studies of erythroid cell development and infectious diseases with erythrocyte involvement. Glycophorins may play a role in preventing cell agglutination.
Development References (7)
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Bain BJ. Leukemia diagnosis: A guide to the FAB classification. 1990.
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Blanchard D, Roux YP-L, Vusio P, Follea G. Characterization of monoclonal antibodies directed to human red blood cell glycophorins A and B. 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; 2002:579-582.
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Gross S, Helm K, Gruntmeir JJ, Stillman WS, Pyatt DW, Irons RD. Characterization and phenotypic analysis of differentiating CD34+ human bone marrow cells in liquid culture. Br J Haematol. 1997; 5(318):326. (Clone-specific: Flow cytometry). View Reference
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Keren DF, Hanson CA, Hurtubise PE. David F. Keren, Curtis A. Hanson, Paul E. Hurtubise., ed. Flow cytometry and clinical diagnosis. Chicago: ASCP Press; 1994:1-676.
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Loken MR, Civin CI, Bigbee WL, Langlois RG, Jensen RH. Coordinate glycosylation and cell surface expression of glycophorin A during normal human erythropoiesis. Blood. 1987; 70(6):1959-1961. (Biology). View Reference
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Nakahata T, Okumura N. Cell surface antigen expression in human erythroid progenitors: erythroid and megakaryocytic markers. Leuk Lymphoma. 1994; 13(5-6):401-409. (Biology). View Reference
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Rogers CE, Bradley MS, Palsson BO, Koller MR. Flow cytometric analysis of human bone marrow perfusion cultures: erythroid development and relationship with burst-forming units-erythroid. Exp Hematol. 1996; 24(5):597-604. (Biology). View Reference
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