Is Recombinant Human LAG-3 His-tag Protein, CF (Catalog # 10056-L3) a monomer or a dimer?

Name: Recombinant Human LAG-3 His-tag Protein, CF
Brand: R&D Systems
SKU: 10056-L3

In house, we observed this 10056-L3 protein as a monomer. In SDS-PAGE, under reducing and non-reducing conditions, the band runs at nearly the same MW.

Recombinant Human LAG-3 His-tag Protein, CF

Catalog # 10056-L3

Product Datasheet / COA / SDS

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Key Product Details

Source:	CHO
Accession #:	P18627
Applications:	Bioactivity

View Full Technical Details for Recombinant Human LAG-3 His-tag Protein, CF

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).

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

	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 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

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

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.

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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).

Item	Value
References	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.
Long Name	Lymphocyte-activation Gene 3
Entrez Gene IDs	3902 (Human); 16768 (Mouse); 297596 (Rat); 102122272 (Cynomolgus Monkey)
Alternate Names	CD223, CD223 antigen, LAG-3, LAG3, Secreted lymphocyte activation gene 3 protein, lymphocyte activating 3, lymphocyte activation gene 3 protein, lymphocyte-activation gene 3, sLAG-3

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FAQs

Is Recombinant Human LAG-3 His-tag Protein, CF (Catalog # 10056-L3) a monomer or a dimer?

In house, we observed this 10056-L3 protein as a monomer. In SDS-PAGE, under reducing and non-reducing conditions, the band runs at nearly the same MW.