Product Information

Contents: Affinity Purified anti-mouse CD223 (LAG3, LAG-3)
Catalog Number: 14-2231
Sizes: 50 ug, 100 ug, 500 ug
Formulation: Phosphate buffer pH 7.2,
150 mM NaCl, 0.09% NaN₃
Storage Conditions: Store at 4°C.
Avoid repeated freeze/thaw cycles.
Clone: eBioC9B7W (C9B7W)
Isotype: Rat IgG1, κ

Staining of 3 day anti-CD3 and anti-CD28-stimulated C57Bl/6 splenocytes with 0.25 μg of Purified Rat IgG1 Iso Cntrl (cat. 14-4301) (open histogram) or 0.25 μg of Purified anti-mouse CD223 (C9B7W) (colored histogram) followed by Biotin Anti-Rat IgG (cat. 13-4813)and SAv-PE (cat. 12-4317). Total viable cells were used for analysis.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
12-2231	Phycoerythrin (PE) anti-mouse CD223 (LAG3, LAG-3)	488	575	FC
14-2231	Affinity Purified anti-mouse CD223 (LAG3, LAG-3)	N/A	N/A	ELISA FA FC IP
16-2231	Functional Grade* Purified anti-mouse CD223 (LAG3, LAG-3)	N/A	N/A	ELISA cap FA FC IP
*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.

Questions? Please consult our answers to frequently asked questions at http://www.ebioscience.com/faq.

Description

The 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 Reported

For 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 Tested

This 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)

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