Preparation And Storage
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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
- 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.
- 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.
- 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.
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- 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 observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
Companion Products
The BRIC 203 monoclonal antibody specifically binds to the Kell blood group antigen, Kp(a-b+) but not Kp(a+b-), also known as the Kell blood group metallo-endopeptidase, KELL, ECE3 and CD238. CD238 is an approximately 115-kDa type II transmembrane glycoprotein that is encoded by the KEL gene. CD238 functions as a zinc endopeptidase that cleaves endothelin-3 into an active form which can function as a vasoconstrictor. CD238 is expressed on erythrocytes, human embryonic stem cell-derived endoderm and on cells from various other tissues, including testis, heart, brain and skeletal muscle. Decreased expression of CD238 is associated with the McLeod phenotype as a consequence of losing XK protein expression. Kell antigen expression can play an important role during blood transfusions and also specific autoimmune diseases related to Kell antigen recognition. During development the BRIC 203 monoclonal antibody was found to detect the CD238 antigen by flow cytometric analysis of human erythrocytes. It reportedly works in immunoprecipitation and agglutination assays.
Development References (4)
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Jung HH, Hergersberg M, Vogt M, et al. McLeod phenotype associated with a XK missense mutation without hematologic, neuromuscular, or cerebral involvement. Transfusion. 2003; 43(7):928-938. (Clone-specific: Flow cytometry). View Reference
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Parsons SF, Gardner B, Anstee DJ. Monoclonal antibodies against Kell glycoprotein: serology, immunochemistry and quantification of antigen sites. Transfus Med. 1993; 3(2):137-142. (Immunogen). View Reference
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Wang P, Rodriguez RT, Wang J, Ghodasara A, Kim SK. Targeting SOX17 in Human Embryonic Stem Cells Creates Unique Strategies for Isolating and Analyzing Developing Endoderm. Cell Stem Cell. 2011; 8:335-346. (Biology). View Reference
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van der Schoot CE, Baardman R, Lighthart P, de Jong I, von dem Borne AEK, de Haas M. CD238 - Kell. 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; 2002:584-585.
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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.