Recombinant Human LAG-3 His-tag Protein, CF Summary
Additional Information |
Analyzed by SEC-MALS |
Details of Functionality |
Measured by its ability to induce TNF-alpha secretion by JAWSII mouse immature dendritic cells. The ED 50 for this effect is 0.2-1.2 μg/mL in the presence of a cross-linking antibody, Mouse Anti-polyHistidine Monoclonal Antibody
(Catalog #
MAB050R). |
Source |
Chinese Hamster Ovary cell line, CHO-derived human LAG-3 protein Leu23-Leu450, with a C-terminal 6-His tag |
Accession # |
|
N-terminal Sequence |
Leu23 |
Protein/Peptide Type |
Recombinant Proteins |
Purity |
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. |
Endotoxin Note |
<0.10 EU per 1 μg of the protein by the LAL method. |
Applications/Dilutions
Dilutions |
|
Theoretical MW |
47 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
SDS-PAGE |
57-63 kDa, reducing conditions
|
Packaging, Storage & Formulations
Storage |
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 3 months, ≤ -20 °C under sterile conditions after reconstitution.
|
Buffer |
Lyophilized from a 0.2 μm filtered solution in PBS. |
Purity |
>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining. |
Reconstitution Instructions |
Reconstitute at 500 μg/mL in PBS. |
Notes
This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.
Alternate Names for Recombinant Human LAG-3 His-tag Protein, CF
Background
LAG-3
(Lymphocyte activation gene-3), designated CD223, is a 70 kDa type I
transmembrane protein that is a member of the immunoglobulin superfamily (IgSF)
(1, 2). LAG-3 shares approximately 20% amino acid sequence homology with
CD4, but has similar structure and binds to MHC class II with higher affinity,
providing negative regulation of T cell receptor signaling (1, 2). Human
LAG-3 cDNA encodes 525 amino acids (aa) that include a 28 aa signal sequence, a
422 aa extracellular domain (ECD) with four Ig-like domains, a
transmembrane region and a highly charged cytoplasmic region. Within the ECD,
human LAG-3 shares 70%, 67%, 76%, and 73% aa sequence identity with mouse, rat,
porcine, and bovine LAG-3, respectively. LAG-3 is expressed on activated CD4
+
and CD8+ T cells, NK cells, and plasmacytoid dendritic cells (pDC),
but not on resting T cells (1-3). LAG-3 on activated CD4
+CD25
+
Treg cells plays a role in their suppressive activity (4). LAG-3
limits the expansion of activated T cells and pDC in response to selected
stimuli (3-5). A soluble 54 kDa form, sLAG-3, can be shed by metalloproteinases
ADAM10 and TACE/ADAM17 (6, 7). While monomeric sLAG-3 itself may be inactive,
shedding allows for normal T cell activation by removing negative regulation
(7). Binding of a homodimerized sLAG-3/Ig fusion protein to MHC class II
molecules induces maturation of immature DC, and secretion of cytokines such as
IFN-gamma and TNF-alpha by type 1 cytotoxic CD8
+ T cells and NK
cells (8, 9). sLAG-3/Ig has been used as a potential adjuvant to stimulate a
cytotoxic anti-cancer immune response (9, 10). In mice, deletion of LAG-3 and
another negative regulator, PD-1, facilitates anti-cancer response but also
blocks self-tolerance and increases susceptibility to autoimmune diseases (11, 12). In humans, antibody-mediated down‑regulation of LAG-3 and PD-1 allows more
effective control of chronic malaria, while in NOD (non‑obese diabetic) mice,
deletion of LAG-3 alone accelerates diabetes (12-14). LAG-3 is an immune checkpoint protein that
modulates T-cell activation and homeostasis and is a promising target for
cancer immunotherapy (15, 16).
- Triebel, F. et al. (1990) J. Exp. Med. 171:1393.
- Baixeras, E. et al. (1992) J. Exp. Med 176:327.
- Workman, C.J. et al. (2004) J. Immunol. 172:5450.
- Huang, C.T. et al. (2004) Immunity 21:503.
- Workman, C.J. et al. (2009) J. Immunol. 182:1885.
- Li, N. et al. (2004) J. Immunol. 173:6806.
- Li, N. et al. (2007) EMBO J. 26:494.
- Andreae, S. et al. (2003) Blood 102:2130.
- Brignone, C. et al. (2007) J. Immunol. 179:4202.
- Brignone, C. et al. (2010) J. Transl. Med. 8:71.
- Woo, S.R. et al. (2011) Cancer Res. 72:917.
- Okazaki, T. et al. (2011) J. Exp. Med. 208:395.
- Bettini, M. et al. (2011) J. Immunol. 187:3493.
- Butler, N.S. et al. (2012) Nat. Immunol. 13:188.
- Durham, N.M. et al. (2014) PLoS One 9:e109080.
- Deng, W.W. et al. (2016) Oncoimmunology 5:e1239005.
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