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Anti-LAG3 monoclonal antibody, recombinant

Catalog #: EG-374

$250.00

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EG-374 Anti-LAG3-M36

Product Name	Anti-LAG3 monoclonal antibody, recombinant
Cat #	EG-374
Size	100 µg
Key features	Rabbit monoclonal antibody produced in 293 cells with serum-free, protein-free medium, purified with protein A column; High batch-to-batch consistency and reproducibility; Long term security of supply;
Clone number	M36
Immunogen	Recombinant extracellular domain of LAG3 that was expressed and purified from 293 cells
Reaction target	Extracellular region of LAG3
Species Cross-Reactivity	Human
Applications	Flow Cytometry; IP; ELISA;
Working concentrations	Flow cytometry and ELISA: 5 µg/ml of final concentration is suggested. IP: 10 μg in 0.3-0.5 mg cell lysates is suggested.
Background	LAG3 (CD223) is a new type of immune check point protein expressed on activated T cells. LAG3 upregulation is required to control overt activation of T cells and prevent the onset of autoimmunity (1). Persistent antigen stimulation results in sustained LAG3 expression, leading to T cell exhaustion. Currently, LAG3 neutralizing antibodies alone or in combination with PD-1/PD-L1 blockade are being rigorously explored as potent immunotherapy strategy to treat solid cancers in clinical trials. Several LAG3 ligands have been proposed, including MHC II, FGL-1, and Galectin-3. Cell surface LAG3 may be cleaved by ADAM10/17 metalloproteinases and released into the blood stream (1). M36 antibody weakly blocks the binding of recombinant LAG3 to MHC II-expressing Raji cells at high concentration. References: (1) Lawrence P Andrews, et al. LAG3 (CD223) as a cancer immunotherapy target. Immunol Rev 2017; 276:80-96.
QC	A) Flow cytometry analysis of cell surface expression of LAG3. Peripheral blood mononuclear cells (PBMC) were activated in vitro with anti-CD3/anti-CD28 beads. Cell surface expression of LAG3 of activated PBMC was detected with the antibody. Activated PBMC (10^6) were incubated with 5 μg antibody in 1 ml PBS buffer containing 5% BSA. Antibody binding was detected with Cy5-conjugated goat-anti-rabbit antibody. Shaded area, isotype control antibody staining; red line, PBMC activated for 1 day; Green line, PBMC activated for 3 days; Blue line, PBMC activated for 7 days. B) Blocking activity of LAG3 monoclonal antibody M36. Recombinant extracellular domain of LAG3 (LAG3ECD-hFc) was shown binding to Raji cells at 10 μg/ml. M36 does not block the binding of LAG3 to Raji cells at 1 and 5 μg/ml, partially blocks at higher concentration of 20 μg/ml.
Concentration	As indicated on the vials
Formulation	80% PBS, pH 7.4; 20% Glycerol; 5% Trehalose; 0.04% Sodium azide as Preservative
Shipping	Blue ice
Storage	-20 to -70 ^oC (long term) or 4 ^oC (one week). Depending on the expected frequency of use, the product can be aliquoted to avoid heavily repeated freezing/thawing cycles.

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