This protein datasheet indicates I need to use a cross-linking antibody, Catalog # MAB050, for biological activity. What is this antibody and is it really necessary?

Name: Recombinant Mouse GITR Ligand/TNFSF18 (aa 47-173) Protein
Brand: R&D Systems
SKU: 2177-GL
Price: 394 USD

The antibody is directed against a 6x histidine repeat and is recommended for use as a cross-linker of proteins with 6x his-tag. Crosslinking is often used for proteins that require receptor trimerization and can result greater biological activity. R&D Systems Quality Control tests the performance of these proteins in the presence of the cross-linking antibody. Therefore, it is necessary to use this antibody when trying to achieve the same level of specific activity described in the datasheet.

Recombinant Mouse GITR Ligand/TNFSF18 (aa 47-173) Protein

R&D Systems, Inc. a Bio-Techne Brand

Catalog # 2177-GL

Citations (3)

Product Datasheet / COA / SDS

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Key Product Details

Source:	NS0
Accession #:	AAO89011
Applications:	Bioactivity

View Full Technical Details for Recombinant Mouse GITR Ligand/TNFSF18 (aa 47-173) Protein

Product Specifications for Recombinant Mouse GITR Ligand/TNFSF18 (aa 47-173) Protein

Source	Mouse myeloma cell line, NS0-derived mouse GITR Ligand/TNFSF18 protein Thr47-Ser173, with an N-terminal 10-His tag
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.
N-terminal sequence Analysis	His
Predicted Molecular Mass	16 kDa
SDS-PAGE	16-26 kDa, reducing conditions
Activity	Measured by its ability to stimulate mouse T cell proliferation in the presence of anti-CD3. The ED₅₀ for this effect is 0.02‑0.3 µg/mL in the presence of 10 µg/mL of a cross-linking antibody, Mouse Anti-polyHistidine Monoclonal Antibody (Catalog # MAB050).

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.

2177-GL

Carrier

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Reconstitution	Reconstitute at 100 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.
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.

2177-GL/CF

Carrier Free

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution	Reconstitute at 100 μg/mL in sterile 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.

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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: GITR Ligand/TNFSF18

GITR Ligand, also known as TNFSF18 and TL6, is an approximately 30 kDa type II transmembrane glycoprotein in the TNF superfamily (1). Mouse GITR Ligand consists of a 23 amino acid (aa) cytoplasmic domain, a 19 aa transmembrane segment, and a 131 aa extracellular domain (ECD) (2 ‑ 4). Within the ECD, mouse GITR Ligand shares 56% and 81% aa sequence identity with human and rat GITR Ligand, respectively. GITR Ligand is expressed on antigen presenting cells, CD4^-CD8^- double negative thymic precursors, vascular endothelial cells, neurons, and in the eye (4 ‑ 11). Its expression is transiently up‑regulated by proinflammatory stimulation (5, 8, 11). The binding of GITR Ligand to GITR on mouse CD25⁺ Treg cells permits the reactivation of T cells from Treg‑induced suppression, although this does not appear to occur in humans (9, 12 ‑ 14). GITR Ligand binding to GITR additionally provides a costimulatory signal to activated CD4⁺ and CD8⁺ T cells and NK cells (6, 15, 16). This interaction also induces reverse signaling in GITR Ligand expressing dendritic cells to suppress cellular activation through the same pathway induced by the immunosuppressant dexamethasone (17). In the brain, GITR Ligand/GITR interactions enhance NGF‑mediated neurite outgrowth from sympathetic neurons (10).

Item	Value
References	Azuma, M. (2010) Crit. Rev. Immunol. 30:547. Yu, K.Y. et al. (2003) Biochem. Biophys. Res. Commun. 310:433. Kim, J.D. et al. (2003) Genes Immun. 4:564. Tone, M. et al. (2003) Proc. Natl. Acad. Sci. 100:15059. Stephens, G.L. et al. (2004) J. Immunol. 173:5008. Hanabuchi, S. et al. (2006) Blood 107:3617. Kamimura, Y. et al. (2009) J. Immunol. 182:2708. Hwang, H. et al. (2010) J. Neurosci. Res. 88:2188. Tuyaerts, S. et al. (2007) J. Leukoc. Biol. 82:93. O’Keefe, G.W. et al. (2008) Nat. Neurosci. 11:135. Kim, B.J. et al. (2004) Invest. Ophthalmol. Vis. Sci. 45:3170. Shimizu, J. et al. (2002) Nat. Immunol. 3:135. Ji, H. et al. (2004) J. Immunol. 172:5823. McHugh, R.S. et al. (2002) Immunity 16:311. Kanamaru, F. et al. (2004) J. Immunol. 172:7306. Ronchetti, S. et al. (2004) Eur. J. Immunol. 34:613. Grohmann, U. et al. (2007) Nat. Med. 13:579.
Long Name	Glucocorticoid Induced TNF Receptor Family Related Gene Ligand
Entrez Gene IDs	8995 (Human); 240873 (Mouse); 364031 (Rat)
Alternate Names	AITRL, AITRLMGC138237, Activation-inducible TNF-related ligand, GITR Ligand, GITRL, GITRLglucocorticoid-induced TNFR-related protein ligand, Glucocorticoid-induced TNF-related ligand, TL6AITR ligand, TNFSF18, hGITRLGITR ligand, tumor necrosis factor (ligand) superfamily, member 18, tumor necrosis factor ligand superfamily member 18

Citations for Recombinant Mouse GITR Ligand/TNFSF18 (aa 47-173) Protein (3)

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Citations are publications that use Bio-Techne products. Selected citations for Recombinant Mouse GITR Ligand/TNFSF18 (aa 47-173) Protein include:

Species: Mouse
Sample Types: Whole Cells
Applications: Bioassay

T Sekiya et al. (2018), Nr4a Receptors Regulate Development and Death of Labile Treg Precursors to Prevent Generation of Pathogenic Self-Reactive Cells Cell Rep, 24(6):1627-1638.e6.
PMID: 30089271
Species: Mouse
Sample Types: Whole Cells
Applications: Bioassay

(2012), Regulation of innate CD8+ T-cell activation mediated by cytokines. Proc Natl Acad Sci U S A, 109(25):9971-6.
PMID: 22665806
Species: Mouse
Sample Types: In Vivo
Applications: In Vivo

Kayamuro H et al. (2009), TNF superfamily member, TL1A, is a potential mucosal vaccine adjuvant. Biochem. Biophys. Res. Commun., 384(3):296-300.
PMID: 19406102

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FAQs for Recombinant Mouse GITR Ligand/TNFSF18 (aa 47-173) Protein

This protein datasheet indicates I need to use a cross-linking antibody, Catalog # MAB050, for biological activity. What is this antibody and is it really necessary?

The antibody is directed against a 6x histidine repeat and is recommended for use as a cross-linker of proteins with 6x his-tag. Crosslinking is often used for proteins that require receptor trimerization and can result greater biological activity. R&D Systems Quality Control tests the performance of these proteins in the presence of the cross-linking antibody. Therefore, it is necessary to use this antibody when trying to achieve the same level of specific activity described in the datasheet.

Recombinant Mouse GITR Ligand/TNFSF18 (aa 47-173) Protein

R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

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Product Specifications for Recombinant Mouse GITR Ligand/TNFSF18 (aa 47-173) Protein

Formulation, Preparation and Storage

Carrier Free

2177-GL

Carrier

2177-GL/CF

Carrier Free

Reconstitution Calculator

Background: GITR Ligand/TNFSF18

Citations for Recombinant Mouse GITR Ligand/TNFSF18 (aa 47-173) Protein (3)

FAQs for Recombinant Mouse GITR Ligand/TNFSF18 (aa 47-173) Protein

Product Documents for Recombinant Mouse GITR Ligand/TNFSF18 (aa 47-173) Protein

Product Datasheets

COA

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SDS

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