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-61 monoclonal antibody specifically recognizes a type II transmembrane glycoprotein, CD26, which is a serine exoprotease identified as dipeptidyl/peptidase IV. Among various biological activities, rat CD26 binds fibronectin and collagen. Rat CD26 is involved in the costimulation of thymocyte proliferation in vitro, particularly the CD4-/CD8- subset, and is developmentally regulated on hematopoietic cells. Although mouse and human CD26 anchor ADA (adenosine deaminase) to cell membranes, rat CD26 does not function as an ADA-binding protein. Rat CD26 is expressed in lung endothelial cells, as well as in various epithelial cells. T cells express lower levels of CD26 than CD4-/CD8- thymocytes. The distribution of CD26 antigen in rat bone marrow cells is similar to that of human CD26. OX-61 monoclonal antibody stains CD4+, CD8+, and Ig+ lymphocytes, and the staining increases upon activation.
Development References (8)
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Bristol LA, Sakaguchi K, Appella E, Doyle D, Takacs L. Thymocyte costimulating antigen is CD26 (dipeptidyl-peptidase IV). Costimulation of granulocyte, macrophage, and T lineage cell proliferation via CD26. J Immunol. 1992; 149(2):367-372. (Biology). View Reference
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Gorrell MD, Wickson J, McCaughan GW. Expression of the rat CD26 antigen (dipeptidyl peptidase IV) on subpopulations of rat lymphocytes. Cell Immunol. 1991; 134(1):205-215. (Clone-specific: Immunohistochemistry, Immunoprecipitation). View Reference
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Hanski C, Huhle T, Gossrau R, Reutter W. Direct evidence for the binding of rat liver DPP IV to collagen in vitro. Exp Cell Res. 1988; 178(1):64-72. (Biology). View Reference
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Iwaki-Egawa S, Watanabe Y, Fujimoto Y. CD26/dipeptidyl peptidase IV does not work as an adenosine deaminase-binding protein in rat cells. Cell Immunol. 1997; 178(2):180-186. (Biology). View Reference
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Johnson RC, Zhu D, Augustin-Voss HG, Pauli BU. Lung endothelial dipeptidyl peptidase IV is an adhesion molecule for lung-metastatic rat breast and prostate carcinoma cells. J Cell Biol. 1993; 121(6):1423-1432. (Biology). View Reference
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Kameoka J, Tanaka T, Nojima Y, Schlossman SF, Morimoto C. Direct association of adenosine deaminase with a T cell activation antigen, CD26. Science. 1993; 261(5120):466-469. (Biology). View Reference
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McCaughan GW, Wickson JE, Creswick PF, Gorrell MD. Identification of the bile canalicular cell surface molecule GP110 as the ectopeptidase dipeptidyl peptidase IV: an analysis by tissue distribution, purification and N-terminal amino acid sequence. Hepatology. 1990; 11(4):534-544. (Immunogen: Immunohistochemistry, Immunoprecipitation, Western blot). View Reference
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Piazza GA, Callanan HM, Mowery J, Hixson DC. Evidence for a role of dipeptidyl peptidase IV in fibronectin-mediated interactions of hepatocytes with extracellular matrix. Biochem J. 1989; 262(1):327-334. (Biology). 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.