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 mCD30.1 monoclonal antibody specifically recognizes CD30. CD30 is also known as Tumor necrosis factor receptor superfamily, member 8 (Tnfrsf8). The CD30 molecule is predominantly expressed by activated T lymphocytes, with its expression peaking at day 4-5 on spleen cells activated with plate-bound anti-CD3e antibody. The mCD30.1 antibody reacts with a majority of CD8+ T cells, as well as some CD4+ T cells in these cultures. Expression of CD30 on activated T lymphocytes is regulated by CD28 and cytokines. Its TNF-superfamily ligand, CD30L or CD153, is also expressed on activated T lymphocytes. By northern blot analysis, mouse Cd30 mRNA is detected in the thymus and in 72-hour pokeweed mitogen- and Con A-activated spleen cells, but not in the lung, brain, kidney, liver, bone marrow, unactivated spleen, or 72-hour LPS-activated splenocytes.1 It has also been reported that CD30 is expressed on naive B lymphocytes, it is not detectable after activation, and it starts to return after 72 hours following activation. Reports suggest that signaling through the CD30 molecule may be important in cytokine production by CD8+ CTL lines and may play a role in the regulation of Th1 and Th2 cytokine secretion by CD4+ and CD8+ T cells. It has also been proposed that CD30 plays an important role in the negative selection of thymocytes and protects against autoimmunity. Members of the TNFR family and their ligands are involved in the induction of diverse biological responses in lymphocytes, including differentiation, proliferation, and cellular death. In humans, CD30 was initially identified in Hodgkin and Reed-Sternberg cells in Hodgkin's disease patients and subsequently was found on neoplastic cells of certain types of non-Hodgkin's lymphomas.
Development References (12)
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Amakawa R, Hakem A, Kundig TM, et al. Impaired negative selection of T cells in Hodgkin's disease antigen CD30-deficient mice. Cell. 1996; 84(4):551-562. (Biology). View Reference
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Bowen MA, Lee RK, Miragliotta G, Nam SY, Podack ER. Structure and expression of murine CD30 and its role in cytokine production. J Immunol. 1996; 156(2):442-449. (Immunogen). View Reference
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Chiarle R, Podda A, Prolla G, Podack ER, Thorbecke GJ, Inghirami G. CD30 overexpression enhances negative selection in the thymus and mediates programmed cell death via a Bcl-2-sensitive pathway. J Immunol. 1999; 163(1):194-205. (Biology). View Reference
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Cosman D. A family of ligands for the TNF receptor superfamily. Stem Cells. 1994; 12(5):440-455. (Biology). View Reference
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DeYoung AL, Duramad O, Winoto A. The TNF receptor family member CD30 is not essential for negative selection. J Immunol. 2000; 165(11):6170-6173. (Biology). View Reference
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Falini B, Pileri S, Pizzolo G, et al. CD30 (Ki-1) molecule: a new cytokine receptor of the tumor necrosis factor receptor superfamily as a tool for diagnosis and immunotherapy. Blood. 1995; 85(1):1-14. (Biology). View Reference
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Gilfillan MC, Noel PJ, Podack ER, Reiner SL, Thompson CB. Expression of the costimulatory receptor CD30 is regulated by both CD28 and cytokines. J Immunol. 1998; 160(5):2180-2187. (Biology). View Reference
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Heath WR, Kurts C, Caminschi I, Carbone FR, Miller JF. CD30 prevents T-cell responses to non-lymphoid tissues. Immunol Rev. 1999; 169:23-29. (Biology). View Reference
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Nakamura T, Lee RK, Nam SY, et al. Reciprocal regulation of CD30 expression on CD4+ T cells by IL-4 and IFN-gamma. J Immunol. 1997; 158(5):2090-2098. (Biology). View Reference
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Shanebeck KD, Maliszewski CR, Kennedy MK, et al. Regulation of murine B cell growth and differentiation by CD30 ligand. Eur J Immunol. 1995; 25(8):2147-2153. (Biology). View Reference
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Shimozato O, Takeda K, Yagita H, Okumura K. Expression of CD30 ligand (CD153) on murine activated T cells. Biochem Biophys Res Commun. 1999; 256(3):519-526. (Biology). View Reference
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Siegmund T, Armitage N, Wicker LS, Peterson LB, Todd JA, Lyons PA. Analysis of the mouse CD30 gene: a candidate for the NOD mouse type 1 diabetes locus Idd9.2. Diabetes. 2000; 49(9):1612-1616. (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.