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.
- 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.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
Companion Products
The OX-38 antibody specifically recognizes the CD4 antigen on most thymocytes, a subpopulation of mature T lymphocytes (ie, MHC class II-restricted T cells, including most T helper cells), monocytes, macrophages, and some dendritic cells. CD4 is an antigen coreceptor on the T-cell surface which interacts with MHC class II molecules on antigen-presenting cells. It participates in T-cell activation through its association with the T-cell receptor complex and protein tyrosine kinase lck. The OX-38 antibody has been reported to bind to the same epitope of CD4 as that recognized by W3/25 mAb, which is a different epitope than that recognized by OX-35 mAb. In vivo blocking of some cell-mediated immune responses by mAb OX-38 has been reported. Injection of OX-38 mAb induces allograft unresponsiveness in rats, with varying results depending on the rat strain used (high or low responder). Furthermore, in vivo depletion of CD4+ lymphocytes has been reported with this antibody.
Development References (9)
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Arima T, Goss JA, Walp LA, Flye MW. Administration of anti-CD4 monoclonal antibody with intrathymic injection of alloantigen results in rat cardiac allograft tolerance. Surgery. 1995; 118(2):265-273. (Clone-specific: Depletion). View Reference
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Bañuls MP, Alvarez A, Ferrero I, Zapata A, Ardavin C. Cell-surface marker analysis of rat thymic dendritic cells. Immunology. 1993; 79(2):298-304. (Biology). View Reference
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Bierer BE, Sleckman BP, Ratnofsky SE, Burakoff SJ. The biologic roles of CD2, CD4, and CD8 in T-cell activation. Annu Rev Immunol. 1989; 7:579-599. (Biology). View Reference
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Janeway CA Jr. The T cell receptor as a multicomponent signalling machine: CD4/CD8 coreceptors and CD45 in T cell activation. Annu Rev Immunol. 1992; 10:645-674. (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: Immunoprecipitation).
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Liu L, Zhang M, Jenkins C, MacPherson GG. Dendritic cell heterogeneity in vivo: two functionally different dendritic cell populations in rat intestinal lymph can be distinguished by CD4 expression. J Immunol. 1998; 161(3):1146-1155. (Biology). View Reference
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Stitz L, Sobbe M, Bilzer T. Preventive effects of early anti-CD4 or anti-CD8 treatment on Borna disease in rats. J Virol. 1992; 66(6):3316-3323. (Clone-specific: Blocking). View Reference
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Suzuki H, Hara MH, Miyahara T, et al. Microchimerism and graft acceptance: IV. Cardiac allograft acceptance following anti-adhesion molecule antibody therapy. Transplant Proc. 1996; 28(4):2058-2060. (Clone-specific: Blocking). View Reference
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Yin D, Fathman CG. Tissue-specific effects of anti-CD4 therapy in induction of allograft unresponsiveness in high and low responder . Transpl Immunol. 1995; 3(3):258-264. (Clone-specific: Blocking). 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.