| 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. |