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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.
Companion Products
The 8F4H7B7 antibody reacts with Vδ 6.3/2 T-cell Receptor (TCR)-bearing T lymphocytes. Originally defined as a member of the Vδ 6 TCR subfamily, it is now proposed that the C57BL-derived Vδ 6.3 is an allelic variant of Vδ 6.2, found in A/J, AKR, BALB/c, C3H/He, and FVB mice. mAb 8F4H7B7 crossreacts with Vδ 6.4 and possibly Vδ 6.6 in DBA/2 mice, and it also detects a subset of γδ TCR-bearing cells in CBA/J and C57L mice. It does not recognize Vδ 4, Vδ 5, Vδ 6.1, or Vδ 6.5 TCR. A subpopulation of thymocytes expressing Vδ 6.3 or Vδ 6.4 TCR (in C57BL/6 or DBA/2 mice, respectively) and low levels of CD90.2 (Thy-1.2) shares functional and phenotypic characteristics with NK-T cells. Similar δV 6.4 TCRexpressing lymphocytes make up significant proportions of the γδ T-cell populations in the liver and spleen of DBA/2 mice. Furthermore, T lymphocytes bearing Vδ 6.3/2 TCR are found in the skin and intestinal epithelium and may represent a unique T-cell subpopulation with a potential for autoimmune reactivity.
Development References (5)
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Azuara V, Grigoriadou K, Lembezat MP, Nagler-Anderson C, Pereira P. Strain-specific TCR repertoire selection of IL-4-producing Thy-1 dull gamma delta thymocytes. Eur J Immunol. 2001; 31(1):205-214. (Immunogen). View Reference
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Azuara V, Lembezat MP, Pereira P. The homogeneity of the TCRdelta repertoire expressed by the Thy-1dull gammadelta T cell population is due to cellular selection. Eur J Immunol. 1998; 28(11):3456-3467. (Biology). View Reference
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Gerber DJ, Azuara V, Levraud JP, Huang SY, Lembezat MP, Pereira P. IL-4-producing gamma delta T cells that express a very restricted TCR repertoire are preferentially localized in liver and spleen. J Immunol. 1999; 163(6):3076-3082. (Biology). View Reference
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Kalataradi H, Eyster CL, Fry A. Allelic differences in TCR gamma-chains alter gamma delta T cell antigen reactivity. J Immunol. 1994; 153(4):1455-1465. (Biology). View Reference
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Nagler-Anderson C, McNair LA, Cradock A. Self-reactive, T cell receptor-gamma delta+, lymphocytes from the intestinal epithelium of weanling mice. J Immunol. 1992; 149(7):2315-2322. (Biology). 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.