Recombinant Human LAG-3 His-tag Protein, CF

R&D Systems, part of Bio-Techne | Catalog # 10056-L3

Analyzed by SEC-MALS
R&D Systems, part of Bio-Techne
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10056-L3-050
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Key Product Details

Source

CHO

Accession #

P18627

Conjugate

Unconjugated

Applications

Bioactivity

View all LAG-3 Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human LAG-3 protein
Leu23-Leu450, with a C-terminal 6-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Leu23

Predicted Molecular Mass

47 kDa

SDS-PAGE

57-63 kDa, reducing conditions

Activity

Measured by its ability to induce TNF-alpha secretion by JAWSII mouse immature dendritic cells.
The ED50 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).

Scientific Data Images for Recombinant Human LAG-3 His-tag Protein, CF

Recombinant Human LAG‑3 His-tag Protein SEC-MALS.

Recombinant human LAG-3 (Catalog # 10056-L3) has a molecular weight (MW) of 75.2 kDa as analyzed by SEC-MALS, suggesting that this protein is a monomer.  MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).
Recombinant Human LAG-3 His-tag Protein Bioactivity

Recombinant Human LAG-3 His-tag Protein Bioactivity

Recombinant Human LAG-3 induces TNF-a secretion in JAWSII mouse immature dendritic cells. The ED50 for this effect is 0.2-1.2 µg/mL in the presence of a cross-linking antibody, Mouse Anti-His Tag Monoclonal Antibody (Catalog # MAB050R).
Recombinant Human LAG-3 His-tag Protein SDS-PAGE

Recombinant Human LAG-3 His-tag Protein SDS-PAGE

2 μg/lane of Recombinant Human LAG‑3 was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 57-63 kDa and 55-60 kDa, respectively.

Formulation, Preparation and Storage

10056-L3
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 500 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & 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.

Background: LAG-3

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).

References

  1. Triebel, F. et al. (1990) J. Exp. Med. 171:1393.
  2. Baixeras, E. et al. (1992) J. Exp. Med 176:327.
  3. Workman, C.J. et al. (2004) J. Immunol. 172:5450.
  4. Huang, C.T. et al. (2004) Immunity 21:503.
  5. Workman, C.J. et al. (2009) J. Immunol. 182:1885.
  6. Li, N. et al. (2004) J. Immunol. 173:6806.
  7. Li, N. et al. (2007) EMBO J. 26:494.
  8. Andreae, S. et al. (2003) Blood 102:2130.
  9. Brignone, C. et al. (2007) J. Immunol. 179:4202.
  10. Brignone, C. et al. (2010) J. Transl. Med. 8:71.
  11. Woo, S.R. et al. (2011) Cancer Res. 72:917.
  12. Okazaki, T. et al. (2011) J. Exp. Med. 208:395.
  13. Bettini, M. et al. (2011) J. Immunol. 187:3493.
  14. Butler, N.S. et al. (2012) Nat. Immunol. 13:188.
  15. Durham, N.M. et al. (2014) PLoS One 9:e109080.
  16. Deng, W.W. et al. (2016) Oncoimmunology 5:e1239005.

Long Name

Lymphocyte-activation Gene 3

Alternate Names

CD223, LAG3

Entrez Gene IDs

3902 (Human); 16768 (Mouse); 297596 (Rat); 102122272 (Cynomolgus Monkey)

Gene Symbol

LAG3

UniProt

P18627

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