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Flow cytometric analysis using BD OptiBuild™ RY703 Rat Anti-Mouse CD326 (EpCAM) antibody (Cat. No. 770044; solid line histogram) on live CD4- and CD8- gated BALB/c Mouse thymocytes, with Autofluorescence Control. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer.
BD OptiBuild™ RY703 Rat Anti-Mouse CD326 (EpCAM)
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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.
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- 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.
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- 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 G8.8 monoclonal antibody recognizes CD326/Ep-CAM (Epithelial Cell Adhesion Molecule), also known as gp40 in the mouse and by a variety of names (including GA733-2, CO17-1A, and EGP) in the human. In the mouse, Ep-CAM is a 40-42 kDa cell-surface type 1 transmembrane glycoprotein expressed on thymic epithelial cells, thymic dendritic cells, immature thymocytes, a small subset of peripheral T lymphocytes, intestinal epithelium, kidney-collecting tubule epithelium, keratinocytes, Langerhans cells and lymph node and splenic dendritic cells. Profiles of Ep-CAM expression on fetal thymocytes and on the CD4[-] CD8[-], CD4[+] CD8[+], CD4[-] CD8[+], and CD4[+] CD8[-] subsets of adult thymocytes have been published. In unrelated studies, mouse Ep-CAM mRNA was detected in tissues containing epithelial cells (kidney, stomach, intestine, lung, and thymus) and in plasma cells and plasmacytomas, but not in heart, muscle, liver, brain, spleen, B lymphomas, or pre-B lymphomas. Ep-CAM is a Ca[2+] independent homophilic adhesion molecule that is proposed to play roles in the development and normal function of epithelial tissues and in the progression of carcinomas.
研发参考 (9)
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Basak S, Speicher D, Eck S, Wunner W. Colorectal carcinoma invasion inhibition by CO17-1A/GA733 antigen and its murine homologue. J Natl Cancer Inst. 1998; 90(9):691-697. (Biology). 查看参考
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Bergsagel PL, Victor-Kobrin C, Timblin CR, Trepel J, Kuehl WM. A murine cDNA encodes a pan-epithelial glycoprotein that is also expressed on plasma cells. J Immunol. 1992; 148(2):590-596. (Biology). 查看参考
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Birebent B, Somasundaram R, Purev E. Anti-idiotypic antibody and recombinant antigen vaccines in colorectal cancer patients. Crit Rev Oncol Hematol. 2001; 39((1-2)):107-113. (Biology). 查看参考
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Borkowski TA, Nelson AJ, Farr AG, Udey MC. Expression of gp40, the murine homologue of human epithelial cell adhesion molecule (Ep-CAM), by murine dendritic cells. Eur J Immunol. 1996; 26(1):110-114. (Clone-specific: Immunohistochemistry). 查看参考
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Farr A, Nelson A, Truex J, Hosier S. Epithelial heterogeneity in the murine thymus: a cell surface glycoprotein expressed by subcapsular and medullary epithelium. J Histochem Cytochem. 1991; 39(5):345-353. (Immunogen: Electron microscopy, Immunohistochemistry, Immunoprecipitation). 查看参考
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Litvinov SV, Balzar M, Winter MJ. Epithelial cell adhesion molecule (Ep-CAM) modulates cell-cell interactions mediated by classic cadherins. J Biol Chem. 1997; 139(5):1337-1348. (Biology). 查看参考
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Nelson AJ, Dunn RJ, Peach R, Aruffo A, Farr AG. The murine homolog of human Ep-CAM, a homotypic adhesion molecule, is expressed by thymocytes and thymic epithelial cells. Eur J Immunol. 1996; 26(2):401-408. (Clone-specific: Electron microscopy, Immunohistochemistry, Immunoprecipitation). 查看参考
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Taguchi N, Hashimoto Y, Naiki M. Abnormal thymic expression of epithelial cell adhesion molecule (EP-CAM) in New Zealand Black (NZB) mice. J Autoimmun. 1999; 13(4):393-400. (Clone-specific: Electron microscopy). 查看参考
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Zutter MM. Gastrointestinal carcinoma antigen GA733: target for immunodestruction and potential modifier of invasiveness and chemoresponsiveness. J Natl Cancer Inst. 1998; 90(9):642-644. (Biology). 查看参考
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