Detection of LAG-3 in Human PBMCs by Flow Cytometry.
CD3+ PHA-treated peripheral blood mononuclear cells (PBMCs) were stained with Mouse Anti-Human CD3 epsilon APC‑conjugated Monoclonal Antibody (Catalog # FAB100A) and either (A) Goat Anti-Human LAG‑3 PE‑conjugated Antigen Affinity-purified Polyclonal Antibody (Catalog # FAB2319P) or (B) Normal Goat IgG Phycoerythrin Control (Catalog # IC108P). View our protocol for Staining Membrane-associated Proteins.
Detection of LAG-3 in HEK293 Human Cell Line Transfected with Human LAG-3 and eGFP by Flow Cytometry.
HEK293 human embryonic kidney cell line transfected with either (A) human LAG-3 or (B) irrelelvant transfectants and eGFP was stained with Goat Anti-Human LAG-3 Phycoerythrin-conjugated Antigen Affinity-purified Polyclonal Antibody (Catalog # FAB2319P). Quadrant markers were set based on control antibody staining (Catalog # IC108P, data not shown). View our protocol for Staining Membrane-associated Proteins.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
12 months from date of receipt, 2 to 8 °C as supplied.
LAG-3 (Lymphocyte activation gene-3), also known as CD223, is a member of the immunoglobulin superfamily (IgSF). The mature LAG-3 protein is a 496 amino acid (aa) membrane protein with a 421 aa extracellular region which contains four IgSF domains, a 21 aa transmembrane region and a 54 aa cytoplasmic region. LAG-3 and CD4 molecules share < 20% aa sequence homology but have a similar structure (1, 2). Both molecules bind to MHC class II. LAG-3 binds to MHC class II with higher affinity compared to CD4. Both LAG-3 and CD4 genes are located on the distal part of the short arm of chromosome 12.
LAG-3 is an activation-induced molecule, expressed on activated T cells and NK cells, but not on resting T cells. Studies using LAG-3 -/- mice have shown significant delay of T cell apoptosis following antigen stimulation and increased size of memory T cells pool following infection (3, 4). It also has been reported that anti-LAG-3 antibodies up-regulate T cell activation by blocking interaction of LAG-3 and MHC class II. The study has demonstrated that LAG-3 is selectively expressed on activated CD4+CD25+ TReg cells and plays a role in their suppressive activity (5). This evidence indicated, unlike the interaction of CD4 with MHC class II that plays a positive role in T cell activation, LAG-3 binds to MHC class II and negatively regulates T cell activation through LAG-3 signaling. On the other hand, studies have shown that binding of LAG-3 to MHC class II molecules on antigen presenting cells induce maturation of dendritic cells and cytokine secretion by monocytes through MHC class II signal transduction (6). Taken together, LAG-3 may have two major functions, it negatively regulates T cells activation through LAG-3 signaling and stimulates antigen presenting cells which express MHC class II.
Triebel, F. et al. (1990) J. Exp. Med. 171:1393.
Baixeras, E. et al. (1992) J. Exp. Med 176:327.
Workman, C.J. and D.A. Vignali (2003) Eur. J. Immunol. 33:970.
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