Recombinant Human LAG-3 Fc Chimera Biotinylated Protein, CF

Catalog #: BT2319 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
BT2319-050

Recombinant Human LAG-3 Fc Chimera Biotinylated Protein Binding Activity

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human LAG-3 Fc Chimera Biotinylated Protein, CF Summary

Product Specifications

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

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

Activity

Measured by its binding ability in a functional ELISA. When Recombinant Human LSECtin/CLEC4G (Catalog # 2947-CL) is coated at 1 μg/mL, 100 uL/well, biotinylated recombinant LAG-3 Fc Chimera (Catalog# BT2319) binds with an ED₅₀ of 0.2-2 μg/mL.

Source

Mouse myeloma cell line, NS0-derived human LAG-3 protein

Human LAG-3 (Leu23-Leu450) Accession # P18627	IEGRMD	Human IgG₁ (Pro100-Lys330)
N-terminus		C-terminus

Accession #

P18627

N-terminal Sequence
Analysis

Leu23

Structure / Form

Disulfide-linked homodimer. Biotinylated via amines.

Predicted Molecular Mass

72.7 kDa

SDS-PAGE

74-91 kDa, reducing conditions

Product Datasheets

BT2319

Product Datasheet

COA

SDS

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.

BT2319

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution	Reconstitute at 200 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 to -70 °C under sterile conditions after reconstitution.

Scientific Data

Binding Activity

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When Recombinant Human LSECtin/CLEC4G (Catalog # 2947-CL) is coated at 1 µg/mL, 100 µL/well, Biotinylated Recombinant Human LAG-3 Fc Chimera Biotinylated (Catalog # BT2319) binds with an ED₅₀ of 0.2-2 µg/mL.

Reconstitution Calculator

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⁺ T_reg 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).

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.

Long Name

Lymphocyte-activation Gene 3

Entrez Gene IDs

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

Alternate Names

CD223 antigen; CD223; LAG3; LAG-3; lymphocyte activating 3; lymphocyte activation gene 3 protein; lymphocyte-activation gene 3; Secreted lymphocyte activation gene 3 protein; sLAG-3