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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.
商品通知
- 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.
配套商品
The 16F11 monoclonal antibody specifically binds to the extracellular domain of Notch2. This Type 1 transmembrane protein is a member of the Notch family that includes Notch1-4. Notch ligands include membrane-bound Jagged1, Jagged2 and Delta-like1, 3 and 4. Notch receptors mediate juxtacrine signals between adjacent cells. Upon ligand binding, Notch2 undergoes proteolytic cleavage that results in the release of the Notch intracellular domain, NICD. NICD translocates to the nucleus where it forms a transcriptional activator complex with the RBP-J DNA binding factor and other transcriptional cofactors. These multimeric complexes regulate the expression of multiple genes including those that orchestrate many facets of embryonic development and the subsequent functioning of cells within multiple organ systems. Notch2 is expressed on bone marrow cells, DN1 and DN2 thymocytes, B cells and activated T cells. Notch2 plays multiple roles in the immune system including the development of marginal zone B cells and the regulation of granzyme B expression by CD8+ T cells. In altered forms, Notch2 has been implicated in cellular transformation or progression in various neoplasms.
研发参考 (4)
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Fiorini E, Merck E, Wilson A, et al. Dynamic regulation of Notch1 and Notch2 surface expression during T cell development and activation revealed by novel monoclonal antibodies. J Immunol. 2009; 183(11):7212-7222. (Immunogen: Flow cytometry, Immunofluorescence). 查看参考
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Maekawa Y, Minato Y, Ishifune C, et al. Notch2 integrates signaling by the transcription factors RBP-J and CREB1 to promote T cell cytotoxicity. Nat Immunol. 2008; 9(10):1140-1147. (Biology). 查看参考
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Radtke F, Fasnacht N, Macdonald HR. Notch signaling in the immune system. Immunity. 2010; 32(1):14-27. (Biology). 查看参考
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Saito T, Chiba S, Ichikawa M, et al. Notch2 is preferentially expressed in mature B cells and indispensable for marginal zone B lineage development. Immunity. 2003; 18(5):675-685. (Biology). 查看参考
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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.