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.
Companion Products
The OX-34 antibody reacts with CD2 (LFA-2), a member of the immunoglobulin superfamily. In the rat, CD2 is expressed on thymocytes, T lymphocytes in spleen and lymph node, dendritic epidermal T cells, splenic macrophages, and NK cells, but not on B cells, most intestinal intraepithelial lymphocytes, or peritoneal and liver macrophages. CD2 can associate with the T-cell receptor complex, and it may function in both intercellular adhesion and signal transduction. In the rat, CD48 and CD59 have been identified as ligands for CD2. OX-34 mAb binds to the extracellular portion of CD2, and it blocks the binding of CD2 to CD48. While OX-34 antibody does not activate T cells, it partially blocks activation by immobilized mAbs to CD3 (clone G4.18) and αβ T-cell receptor (clone R73), and it partially inhibits allogeneic mixed lymphocyte reactions. Moreover, in vivo administration of OX-34 antibody depletes peripheral T cells and prevents allograft rejection.
Development References (11)
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Beyers AD, Spruyt LL, Williams AF. Molecular associations between the T-lymphocyte antigen receptor complex and the surface antigens CD2, CD4, or CD8 and . Proc Natl Acad Sci U S A. 1992; 89(7):2945-2949. (Biology). View Reference
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Brown MH, Preston S, Barclay AN. A sensitive assay for detecting low-affinity interactions at the cell surface reveals no additional ligands for the adhesion pair rat CD2 and CD48. Eur J Immunol. 1995; 25(12):3222-3228. (Biology). View Reference
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Clark SJ, Law DA, Paterson DJ, Puklavec M, Williams AF. Activation of rat T lymphocytes by anti-CD2 monoclonal antibodies. J Exp Med. 1988; 167(6):1861-1872. (Biology). View Reference
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Elbe A, Kilgus O, Hünig T, and Stingl G. T-cell receptor diversity in dendritic epidermal T cells in the rat. J Invest Dermatol. 1993; 102:74-79. (Biology). View Reference
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Fangmann J, Schwinzer R, Wonigeit K. Unusual phenotype of intestinal intraepithelial lymphocytes in the rat: predominance of T cell receptor alpha/beta+/CD2- cells and high expression of the RT6 alloantigen. Eur J Immunol. 1991; 21(3):753-760. (Biology). View Reference
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Hirahara H, Tsuchida M, Watanabe T, et al. Long-term survival of cardiac allografts in rats treated before and after surgery with monoclonal antibody to CD2. Transplantation. 1995; 59(1):85-90. (Biology). View Reference
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Jefferies WA, Green JR, Williams AF. Authentic T helper CD4 (W3/25) antigen on rat peritoneal macrophages.. J Exp Med. 1985; 162:117-127. (Immunogen).
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Liversidge J, Dawson R, Hoey S, McKay D, Grabowski P, Forrester JV. CD59 and CD48 expressed by rat retinal pigment epithelial cells are major ligands for the CD2-mediated alternative pathway of T cell activation. J Immunol. 1996; 156(10):3696-3703. (Biology). View Reference
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Williams AF, Barclay AN, Clark SJ, Paterson DJ, Willis AC. Similarities in sequences and cellular expression between rat CD2 and CD4 antigens. J Exp Med. 1987; 165(2):368-380. (Biology). View Reference
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van den Brink MR, Hunt LE, Hiserodt JC. In vivo treatment with monoclonal antibody 3.2.3 selectively eliminates natural killer cells in rats. J Exp Med. 1990; 171(1):197-210. (Biology). View Reference
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van der Merwe PA, Brown MH, Davis SJ, Barclay AN. Affinity and kinetic analysis of the interaction of the cell adhesion molecules rat CD2 and CD48. EMBO J. 1993; 12(3):4945-4954. (Biology). 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.