Recombinant Rat TIM-3 Fc Chimera Protein, CF

Name: Recombinant Rat TIM-3 Fc Chimera Protein, CF
Brand: R&D Systems
SKU: 10489-TM

Catalog # 10489-TM

Product Datasheet / COA / SDS

Order Information

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Key Product Details

Source:	NS0
Accession #:	P0C0K5.2
Structure / Form:	Disulfide-linked homodimer
Applications:	Bioactivity

View Full Technical Details for Recombinant Rat TIM-3 Fc Chimera Protein, CF

Product Specifications

Source

Mouse myeloma cell line, NS0-derived rat TIM-3 protein

Rat TIM-3 (Leu22-Ala194) Accession # P0C0K5.2	IEGRMD	Mouse IgG₁ (Pro100-Lys330)
N-terminus		C-terminus

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

Leu22

Predicted Molecular Mass

46 kDa

SDS-PAGE

60-75 kDa, under reducing conditions

Activity

Measured by its binding ability in a functional ELISA. When Recombinant Human Galectin-9 Protein (Catalog # 2045-GA) is immobilized at 1.0 µg/mL (100 µL/ well), Recombinant Rat TIM‑3 Fc Chimera (Catalog # 10489-TM) that produces a 50% optimal binding response is found to be 20-120 ng/mL

Recombinant Rat TIM-3 Fc Chimera Protein, CF Scientific Data Examples

	Recombinant Rat TIM-3 Fc Chimera Protein Bioactivity When Recombinant Human Galectin-9 Protein (2045-GA) is immobilized 1.0 μg/mL (100 μL/well), Recombinant Rat TIM-3 Fc Chimera (10489-TM) that produces a 50% optimal binding response is found to be 20-120 ng/mL.
	Recombinant Rat TIM-3 Fc Chimera Protein SDS-PAGE 2 μg/lane of Recombinant Rat TIM-3 Fc Chimera Protein (10489-TM) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 60-75 kDa and 120-150 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.

10489-TM

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution	Reconstitute at 250 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.

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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Desired Reconstitution Concentration

Background: TIM-3

T cell immunoglobulin and mucin domain-3 (TIM-3) also known as HAVCR2, is a member of the TIM family of immune regulating molecules. TIMs are type I transmembrane glycoproteins with one Ig-like V-type domain, a Ser/Thr-rich mucin stalk region (1, 2). While lacking a specific immunoreceptor tyrosine-based inhibition motif (ITIM), the cytoplasmic domain of TIM-3 contains a conserved region of five tyrosine residues important for downstream signaling (3). A soluble form of TIM-3 lacking the mucin stalk and transmembrane domains is formed as either a result of alternative splicing or metalloproteinase-dependent cleavage. Within the ECD, mature rat TIM-3 shares 60% and 69% amino acid sequence identity with human and mouse TIM-3, respectively. TIM-3 is up‑regulated on several populations of activated myeloid cells (macrophage, monocyte, dendritic cell, microglia, mast cell) and T cells (Th1, CD8+, NK, Treg) (3-10). Its binding to Galectin-9 induces a range of immunosuppressive functions which enhance immune tolerance and inhibit anti-tumor immunity (3,11). TIM-3 ligation attenuates CD8+ and Th1 cell responses (11-13) and promotes the activity of Treg and myeloid derived suppressor cells (8, 11, 13, 14). In addition, dendritic cell-expressed TIM-3 dampens inflammation by enabling the phagocytosis of apoptotic cells and the cross-presentation of apoptotic cell antigens (4). It also binds the alarmin HMGB1, thereby preventing the activation of TLRs in response to released tumor cell DNA (7). TIM-3 interactions with Galectin-9 can alternatively trigger immune stimulatory effects, such as the coactivation of NK cell cytotoxicity (10).

Item	Value
References	Sakuishi, K. et al. (2011) Trends Immunol. 32:345. Anderson, A.C. (2012) Curr. Opin. Immunol. 24:213. Wolf, Y. et al. (2020) Nature Reviews Immunology 20:173. Nakayama, M. et al. (2009) Blood 113:3821. Anderson, A.C. et al. (2007) Science 318:1141. Wiener, Z. et al. (2007) J. Invest. Dermatol. 127:906. Chiba, S. et al. (2012) Nat. Immunol. 13:832. Sanchez-Fueyo, A. et al. (2003) Nat. Immunol. 4:1093. Ndhlovu, L.C. et al. (2012) Blood 119:3734. Gleason, M.K. et al. (2012) Blood 119:3064. Zhu, C. et al. (2005) Nat. Immunol. 6:1245. Sakhdari, A. et al. (2012) PLoS ONE 7:e40146. Sabatos, C.A. et al. (2003) Nat. Immunol. 4:1102. Dardalhon, V. et al. (2010) J. Immunol. 185:1383.
Long Name	T Cell Immunoglobulin Mucin-3
Entrez Gene IDs	84868 (Human); 171285 (Mouse); 363578 (Rat); 479318 (Canine); 102141722 (Cynomolgus Monkey)
Alternate Names	CD366, FLJ14428, HAVCR2, HAVcr-2, KIM-3, SPTCL, T cell immunoglobulin mucin 3, T cell immunoglobulin mucin-3, T-cell immunoglobulin and mucin domain-containing protein 3, TIM 3, TIM-3, TIM3, TIM3 T-cell membrane protein 3, TIMD-3, TIMD3, TIMD3KIM-3, hepatitis A virus cellular receptor 2, kidney injury molecule-3

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