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Questions? Please consult our answers to frequently asked questions at http://www.ebioscience.com/faq. DescriptionThe eBioC9B7W monoclonal antibody recognizes mouse CD223 (LAG-3, LAG3) protein expressed by activated αβ-TCR bearing T cells, a subset of γδ-TCR bearing T cells and a subset of NK cells. CD223 is a 70 kDa type I transmembrane protein with a structure that is similar to CD4. However, a soluble form of human CD223 has been detected by ELISA in human serum, and data suggest that mouse CD223 is proteolytically cleaved in the D4 domain. This results in a 54 kDa fragment containing all the extracellular domains, and a 16 kDa fragment containing the intracellular and transmembrane domains. The 54 kDa can remain membrane-associated or be released as soluble CD223. CD223 binds to MHC class II with higher affinity than CD4, and it is thought that this interaction is involved in the negative regulation of T-cell activation and homeostatic proliferation. Furthermore, CD223 is expressed by CD4+CD25+ regulatory T cells, and it has been suggested that CD223 may be involved in their regulatory function. Applications ReportedFor research use only, not for diagnostic or therapeutic use. This eBioC9B7W (C9B7W) antibody has been reported for use in flow cytometric analysis, immunoprecipitation, and ELISA. (Please use Functional Grade purified eBioC9B7W (C9B7W), cat. 16-2231, in functional assays.) Applications TestedThis eBioC9B7W (C9B7W) antibody has been tested by flow cytometric analysis of anti-CD3 and anti-CD28-activated mouse splenocytes. This can be used at less than or equal to 0.5 μg per million cells in a 100 μl total staining volume. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. This eBioC9B7W (C9B7W) antibody has also been tested by ELISA analysis of supernatant produced from mouse CD223-transfected 3A9 cells. This can be used in ELISA at 2 μg/ml for capture of the secreted form of CD223, together with the polyclonal anti-mouse CD223 (cat. 24-2232) as the detection antibody. References
Baixeras E, Huard B, Miossec C, Jitsukawa S, Martin M, Hercend T, Auffray C, Triebel F, Piatier-Tonneau D. Characterization of the lymphocyte activation gene 3-encoded protein. A new ligand for human leukocyte antigen class II antigens. J Exp Med. 1992 Aug 1;176(2):327-37. Workman CJ, Rice DS, Dugger KJ, Kurschner C, Vignali DA. Phenotypic analysis of the murine CD4-related glycoprotein, CD223 (LAG-3). Eur J Immunol. 2002 Aug;32(8):2255-63. (C9B7W, FC, FA, PubMed) Workman CJ, Vignali DA. Negative regulation of T cell homeostasis by lymphocyte activation gene-3 (CD223). J Immunol. 2005 Jan 15;174(2):688-95. (C9B7W, FA, PubMed) Li N, Workman CJ, Martin SM, Vignali DA. Biochemical analysis of the regulatory T cell protein lymphocyte activation gene-3 (LAG-3; CD223). J Immunol. 2004 Dec 1;173(11):6806-12. (C9B7W, IP, PubMed) Huang CT, Workman CJ, Flies D, Pan X, Marson AL, Zhou G, Hipkiss EL, Ravi S, Kowalski J, Levitsky HI, Powell JD, Pardoll DM, Drake CG, Vignali DA. Role of LAG-3 in regulatory T cells. Immunity. 2004 Oct;21(4):503-13. (C9B7W, FC, FA, PubMed) Related ProductsCat. 00-0000 eBioscience 10X ELISA Coating Buffer Cat. 24-2232 Rabbit anti-mouse CD223 (LAG3, LAG-3) polyclonal serum Cat. 44-2404 Nunc MaxiSorp™ flat-bottom 96 well plate Cat. 18-4100 Avidin Horse Radish Peroxidase (AV-HRP) Cat. 00-4202 eBioscience ELISA Diluent Solution, 5X Cat. 14-4301 Affinity Purified Rat IgG1 Isotype Control Cat. 11-4317 Streptavidin-FITC (Fluorescein isothiocyanate) Cat. 12-4317 Streptavidin-PE (Phycoerythrin) Cat. 17-4317 Streptavidin Allophycocyanin (SA-APC) Cat. 11-4811 FITC Anti-Rat IgG Cat. 13-4813 Biotin Anti-Rat IgG (clone Polyclonal) |
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