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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
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- 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.
Companion Products
The BB7.1 monoclonal antibody specifically recognizes the Human Leukocyte Antigen-B7 (HLA-B7) alpha subunit. The HLA-B7 alpha chain can noncovalently heterodimerize with β2-microglobulin to form a Major Histocompatibility Complex (MHC) class I B7 antigen that is involved in antigen presentation to CD8+ T cells or might serve as an NK cell receptor ligand. HLA-B7 is a serotype that identifies the more common HLA-B (HLA-B*07) encoded gene products which are expressed on nearly all nucleated cells of HLA-B7-positive individuals. B7 is a risk factor for cervical cancer, sarcoidosis, and some spondylarthropathies. The BB7.1 antibody can be used to distinguish false HLA-B27-positve staining when used in conjunction with some HLA-B27 antibodies that crossreact with HLA-B7. Although highly specific, the BB7.1 antibody can reportedly crossreact with HLA-B42 and HLA-B81 antigens.
Development References (4)
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Brodsky FM, Parham P, Barnstable CJ, Crumpton MJ, Bodmer WF. Monoclonal antibodies for analysis of the HLA system. Immunol Rev. 1979; 47:3-6. (Immunogen: Immunoaffinity chromatography, Radioimmunoassay). View Reference
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Hilton HG, Parham P. Direct binding to antigen-coated beads refines the specificity and cross-reactivity of four monoclonal antibodies that recognize polymorphic epitopes of HLA class I molecules.. Tissue Antigens. 2013; 81(4):212-20. (Clone-specific: Cytometric Bead Array). View Reference
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Parham P. Purification of immunologically active HLA-A and -B antigens by a series of monoclonal antibody columns.. J Biol Chem. 1979; 254(18):8709-12. (Clone-specific: Immunoaffinity chromatography, Radioimmunoassay). View Reference
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Reches A, Nachmani D, Berhani O, et al. HNRNPR Regulates the Expression of Classical and Nonclassical MHC Class I Proteins.. J Immunol. 2016; 196(12):4967-76. (Clone-specific: Flow cytometry). View Reference
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.