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推荐的实验流程
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.
- 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.
配套商品
The 34-5-3 antibody reacts with the β chain of the I-A[d] MHC class II alloantigen. It cross-reacts with I-A[b] and with cells from mice of the H-2[p] and H-2[q] haplotypes. Reactivity with other haplotypes (e.g., a, f, k, r, s) has not been observed. The strain distribution of the antigen recognized by this reagent is similar to that of anti-I-A[b] mAb 25-9-17 (Cat. Nos. 553603, 553604, and 553605).
研发参考 (6)
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Bishop GA, Frelinger JA. Haplotype-specific differences in signaling by transfected class II molecules to a Ly-1+ B-cell clone. Proc Natl Acad Sci U S A. 1989; 86(15):5933-5937. (Clone-specific: Stimulation). 查看参考
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Cohn LE, Glimcher LH, Waldmann RA, et al. Identification of functional regions on the I-Ab molecule by site-directed mutagenesis. Proc Natl Acad Sci U S A. 1986; 83(3):747-751. (Biology). 查看参考
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Landias D, Beck BN, Buerstedde JM, et al.. The assignment of chain specificities for anti-Ia monoclonal antibodies using L cell transfectants. J Immunol. 1986; 137(9):3002-3005. (Biology). 查看参考
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Ozato K, Mayer NM, Sachs DH. Monoclonal antibodies to mouse major histocompatibility complex antigens.. Transplantation. 1982; 34(3):113-20. (Immunogen: Cytotoxicity). 查看参考
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Ozato K, Sachs DH. Monoclonal antibodies to mouse MHC antigens. III. Hybridoma antibodies reacting to antigens of the H-2b haplotype reveal genetic control of isotype expression. J Immunol. 1981; 126(1):317-321. (Biology). 查看参考
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Regnier-Vigouroux A, Blanc D, Pont S, Marchetto S, Pierres M. Accessory molecules and T cell activation. I. Antigen receptor avidity differentially influences T cell sensitivity to inhibition by monoclonal antibodies to LFA-1 and L3T4. J Immunol. 1986; 16(11):1385-1390. (Clone-specific: Blocking). 查看参考
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