Mouse GITR Ligand/TNFSF18 Antibody

Catalog #: MAB21773 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB21773-100
MAB21773-SP

Detection of GITR Ligand/TNFSF18 in NS0 Cell Line Transfected with Mouse GITR Ligand/TNFSF18 by Flow Cytometry.

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Mouse GITR Ligand/TNFSF18 Antibody Summary

Species Reactivity

Mouse

Specificity

Detects mouse GITR Ligand/TNFSF18 in direct ELISAs.

Source

Monoclonal Rat IgG₁ Clone # 994529

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse GITR Ligand/TNFSF18
Thr47-Ser173
Accession # Q7TS55

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Endotoxin Level

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

Label

Unconjugated

Applications

Recommended Concentration

Sample

Flow Cytometry

0.25 µg/10⁶ cells

See below

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Neutralization

Measured by its ability to neutralize GITR Ligand/TNFSF18-induced proliferation in mouse CD4+ T cells. The Neutralization Dose (ND₅₀) is typically 1.2-12 µg/mL in the presence of 2 µg/mL Recombinant Mouse GITR Ligand/TNFSF18, 0.5 µg/mL Mouse CD3 Monoclonal Antibody, and 10 µg/mL of a cross-linking antibody, Anti-His Tag Monoclonal Antibody.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Flow Cytometry

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Detection of GITR Ligand/TNFSF18 in NS0 Cell Line Transfected with Mouse GITR Ligand/TNFSF18 by Flow Cytometry. NS0 cell line transfected with either mouse GITR Ligand/TNFRSF18 (filled histogram) or irrelevant protein (open histogram) was stained with Rat Anti-Mouse GITR Ligand/TNFRSF18 Monoclonal Antibody (Catalog # MAB21773) followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0113). View our protocol for Staining Membrane-associated Proteins.

Neutralization

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Cell Proliferation Induced by GITR Ligand/TNFSF18 and Neutralization by Mouse GITR Ligand/TNFSF18 Antibody. In the presence of Rat Anti-Mouse CD3 Monoclonal Anti-body (0.5 µg/mL, MAB4841) and a cross-linking antibody, Mouse Anti-His Tag Monoclonal Antibody (10 µg/mL, MAB050), Recombinant Mouse GITR Ligand/TNFSF18 (2177-GL) stimulates proliferation in mouse CD4+T cells in a dose-dependent manner (orange line), as measured by Resazurin (AR002). Under these conditions, proliferation elicited by Recombinant Mouse GITR Ligand/TNFSF18 (2 µg/mL) is neutralized (green line) by increasing concentrations of Rat Anti-Mouse GITR Ligand/ TNFSF18 Monoclonal Antibody (Catalog # MAB21773). The ND₅₀ is typically 1.2-12 µg/mL.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: GITR Ligand/TNFSF18

Glucocorticoid-induced TNF receptor superfamily-related protein ligand (GITRL) is a member of the TNF superfamily (TNFSF) and has been designated TNFSF18. Mouse GITRL cDNA encodes a 173 amino acid (aa) type II membrane protein with a C-terminal extracellular domain of 131 aa, an N-terminal cytoplasmic domain of 23 aa and a transmembrane domain of 19 aa. It shares approximately 60% aa sequence identity with human GITRL (2). Mouse GITRL is expressed at high levels in macrophages, dendritic cells and B cells. The expression is transiently upregulated by LPS stimulation. GITRL binds to the type I transmembrane protein GITR/TNFRSF18, which is a member of the TNF receptor superfamily that is predominantly expressed on CD25⁺ regulatory CD4⁺ T cells (Treg). GITR is also expressed on naïve CD4⁺ CD25^-T cells, where its expression is upregulated after antigen-driven activation (1, 2). Ligation of GITR has been found to induce nuclear factor (NF)-kappa B activation via TNF receptor-associated factor 2. GITRL provides costimulatory signals for activated CD4⁺ CD25^-T cells to enhance cell proliferation and augment cytokine production. On CD4⁺ CD25⁺ Treg cells, GITRL also provides costimulatory signals to induce proliferation, setting Treg cells in an active/hyperproliferactive state that reverses the suppressive function of Treg cells. GITRL-GITR ligation provides important costimulatory signals that play important roles in modulating diverse T cell functions (1-4).

References

Tone, M. et al. (2003) Proc. Natl. Acad. Sci. USA 100:15059.
Ji, H. et al. (2004) J. Immunology 172:5823.
Kanamaru, F. et al. (2004) J. Immunology 172:7306.
Ronchetti, S. et al. (2004) Eur. J. Immunology 34:613.

Long Name

Glucocorticoid Induced TNF Receptor Family Related Gene Ligand

Entrez Gene IDs

8995 (Human); 240873 (Mouse); 364031 (Rat)

Alternate Names

Activation-inducible TNF-related ligand; AITRL; AITRLMGC138237; GITR Ligand; GITRL; GITRLglucocorticoid-induced TNFR-related protein ligand; Glucocorticoid-induced TNF-related ligand; hGITRLGITR ligand; TL6AITR ligand; TNFSF18; tumor necrosis factor (ligand) superfamily, member 18; tumor necrosis factor ligand superfamily member 18