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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.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
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The C15 monoclonal antibody specifically recognizes the variable alpha 24 region of the α subunit of the human αβ T cell receptor for antigen (TCR Vα24). TCR Vα24 combines with joining region 18 (Jα18) to form Vα24-Jα18 (Arden nomenclature, or TRAV10-TRAJ18 in the IMGT nomenclature). TCR Vα24 can pair with different TCR Vβ region containing TCR β chains. The paired expression of TCR Vα24 with semivariant TCR Vβ11 identifies CD3+TCR αβ+ positive thymocytes and from ~ 0.01-0.92% of peripheral blood CD3+ TCR αβ+ T cells known as invariant NKT (iNKT) cells or Type I NKT cells. iNKT cells can be divided into three subpopulations which are either CD4+, CD8+, or CD4-CD8- double negative. Due to their innate-like responsiveness, tissue resident iNKT cells are capable of rapid effector responses to microbial or self-lipid antigens presented by the cell surface MHC class Ib molecule, CD1d. iNKT cells recognize and respond to α-galactosylceramide (α-GalCer) and various self-lipids such as sphingolipids and diacylglycerols and microbial lipids presented by CD1d. iNKT cells can produce high levels of cytokines, including IL-4 and IFN-γ, and act as cytotoxic T cells against CD1d+ target cells following stimulation. The C15 antibody is useful for multiparameter analyses designed to study the nature of TCR Vα24-positive T cells and T cell clones. When combined with other TCR V region-specific antibodies, C15 is helpful for characterizing TCR Vα repertoires expressed by T cell populations collected from blood, tissues or other sources in health and disease models including inflammation, infectious diseases, autoimmunity, and tumors. The C15 antibody reportedly stimulates the proliferation of TCR Vα24-positive T cells.
Development References (10)
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Arden B, Clark SP, Kabelitz D, Mak TW. Human T-cell receptor variable gene segment families.. Immunogenetics. 1995; 42(6):455-500. (Biology). View Reference
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Dellabona P, Padovan E, Casorati G, Brockhaus M, Lanzavecchia A. An invariant V alpha 24-J alpha Q/V beta 11 T cell receptor is expressed in all individuals by clonally expanded CD4-8- T cells.. J Exp Med. 1994; 180(3):1171-6. (Immunogen: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Elliott JI. Selection of dual Valpha T cells.. Eur J Immunol. 1998; 28(7):2115-23. (Clone-specific: Flow cytometry). View Reference
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Godfrey DI, Uldrich AP, McCluskey J, Rossjohn J, Moody DB. The burgeoning family of unconventional T cells.. Nat Immunol. 2015; 16(11):1114-23. (Biology). View Reference
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Kent SC, Hafler DA, Strominger JL, Wilson SB. Noncanonical Valpha24JalphaQ T cells with conservative alpha chain CDR3 region amino acid substitutions are restricted by CD1d.. Hum Immunol. 1999; 60(11):1080-9. (Clone-specific: Flow cytometry). View Reference
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Montoya CJ, Pollard D, Martinson J, et al. Characterization of human invariant natural killer T subsets in health and disease using a novel invariant natural killer T cell-clonotypic monoclonal antibody, 6B11.. Immunology. 2007; 122(1):1-14. (Clone-specific: Flow cytometry). View Reference
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Padovan E, Casorati G, Dellabona P, Meyer S, Brockhaus M, Lanzavecchia A. Expression of two T cell receptor alpha chains: dual receptor T cells.. Science. 1993; 262(5132):422-4. (Immunogen: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Prell C, Konstantopoulos N, Heinzelmann B, et al. Frequency of Valpha24+CD161+ natural killer T cells and invariant TCRAV24-AJ18 transcripts in atopic and non-atopic individuals.. Immunobiology. 2003; 208(4):367-80. (Clone-specific: Flow cytometry). View Reference
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Scaviner D, Lefranc MP. The human T cell receptor alpha variable (TRAV) genes.. Exp Clin Immunogenet. 2000; 17(2):83-96. (Biology). View Reference
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Vyth-Dreese FA, Sein J, van de Kasteele W, et al. Lack of anti-tumour reactivity despite enhanced numbers of circulating natural killer T cells in two patients with metastatic renal cell carcinoma.. Clin Exp Immunol. 2010; 162(3):447-59. (Clone-specific: Flow cytometry). View Reference
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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.