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.
- 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.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- 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.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
- 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 1082C5 monoclonal antibody (also known as clone 108-2C5) recognizes a shared antigenic determinant present on the extracellular region of Human Leukocyte Antigen-A (HLA-A) heavy chains encoded by a limited number of different HLA-A alleles (HLA-A2, -A3, -A28, -A29, -A30, -A31 and -A33). HLA-A antigens belong to the major histocompatibility complex (MHC) of class I antigens along with HLA-B and HLA-C antigens. HLA class I molecules are heterodimers comprised of an ~40-45 kDa, highly polymorphic transmembrane α heavy chain that is a type I glycoprotein which is noncovalently-associated with an invariant β2-microglobulin (β2m) light chain. The N-terminal extracellular region of the HLA class I heavy chain is comprised of three domains (α1, α2, and α3). The α1 and α2 domains form a closed antigen-binding groove that accommodates 8-10 aa-peptide antigens. β2m non-covalently associates with the α3 heavy chain domain and promotes HLA class I stability. The intralocus HLA-A determinant recognized by the 108-2C5 antibody reportedly involves amino acids residues 76-80 of the HLA-A α1 domain. HLA-A antigens are normally expressed on all nucleated cells. These molecules play central roles in the MHC class I-restricted presentation and cross-presentation of antigens and the regulation of NK and T cell-mediated cytotoxicity that are involved in immune responses to pathogens and tumors as well as tissue allotransplantation.
Development References (6)
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Bardi MS, Jarduli LR, Jorge AJ, et al. HLA-A, B and DRB1 allele and haplotype frequencies in volunteer bone marrow donors from the north of Parana State.. Rev Bras Hematol Hemoter. 2012; 34(1):25-30. (Biology). View Reference
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Guerreiro-Cacais AO, Uzunel M, Levitskaya J, Levitsky V. Inhibition of heavy chain and beta2-microglobulin synthesis as a mechanism of major histocompatibility complex class I downregulation during Epstein-Barr virus replication.. J Virol. 2007; 81(3):1390-400. (Clone-specific: Flow cytometry). View Reference
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Hühn MH, Hultcrantz M, Lind K, Ljunggren HG, Malmberg KJ, Flodström-Tullberg M. IFN-gamma production dominates the early human natural killer cell response to Coxsackievirus infection.. Cell Microbiol. 2008; 10(2):426-36. (Clone-specific: Flow cytometry). View Reference
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Llano M, Gumá M, Ortega M, Angulo A, López-Botet M. Differential effects of US2, US6 and US11 human cytomegalovirus proteins on HLA class Ia and HLA-E expression: impact on target susceptibility to NK cell subsets.. Eur J Immunol. 2003; 33(10):2744-54. (Clone-specific: Flow cytometry). View Reference
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Lozano F, Santos-Aguado J, Borche L, et al. Identification of the amino acid residues defining an intralocus determinant in the alpha 1 domain of HLA-A molecules.. Immunogenetics. 1989; 30(1):50-3. (Clone-specific: Blocking, Flow cytometry, Immunoprecipitation, Radioimmunoassay). View Reference
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Ryschich E, Cebotari O, Fabian OV, et al. Loss of heterozygosity in the HLA class I region in human pancreatic cancer.. Tissue Antigens. 2004; 64(6):696-702. (Clone-specific: Immunohistochemistry). View Reference
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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.