Purified Recombinant Human GITR (Accession # Q9Y5U5)
<0.1 EU/µg as determined by the LAL method
This monoclonal antibody has been 0.2 µm filtered and lyophilized from modified Dulbecco's phosphate buffered saline (1X PBS) pH 7.2 - 7.3 containing 5.0% w/v trehalose with no calcium, magnesium or preservatives present.
Storage and Handling
The lyophilized antibody can be stored desiccated at -20°C to -70°C for up to twelve months. The reconstituted antibody can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted antibody, aseptically aliquot into working volumes and store at -20°C to -70°C in a manual defrost freezer. Avoid repeated freeze thaw cycles. No detectable loss of activity was observed after six months.
Country of Origin
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Applications and Recommended Usage?
Quality Tested by Leinco
ELISA Sandwich: This antibody is useful as the capture antibody in a sandwich ELISA. The suggested coating concentration is 2-8 µg/ml. A suitable detection antibody is PN:G695 at a concentration of approximately 0.1-0.4 µg/ml. A suggested standard for this assay is PN:G652.
Flow Cytometry: It is recommended to use the indirect method for signal enhancement when enumerating cells expressing GITR. A suggested method would be to stain cells expressing GITR with 0.25 µg per 1 - 2.5 x 106 cells in a total staining volume of ≤200 µl followed by PN:M1189.
Western Blotting: To detect Human GITR this monoclonal antibody can be used at a concentration of 1 µg/ml. This monoclonal antibody should be used in conjunction with compatible second-step reagents such as PN:M114 and a chromogenic substrate such as PN:T343. The detection limit for Human GITR is 25 ng/lane under either reducing or non-reducing conditions. The sensitivity of detection may increase up to 50 fold when a chemiluminescent substrate is used. A suitable Western blotting control is PN:G652.
Other Applications Reported In Literature ?
Neutralization: This antibody is useful for neutralization of Human GITR bioactivity. The antibody dose required to neutralize 50% (ND50) of the biological activity of Human GITR (at 10 ng/ml) is 1-2 µg/ml.
Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change.
Mouse Anti-Human Glucocorticoid Induced Tumor Necrosis Factor Receptor (GITR) (Clone 110416) recognizes an epitope on Human GITR. This monoclonal antibody was purified using multi-step affinity chromatography methods such as Protein A or G depending on the species and isotype.
GITR (glucocorticoid-induced tumor necrosis factor receptor) also known as TNFRSF18, is a surface receptor molecule that has been shown to be involved in inhibiting the suppressive activity of T-regulatory cells and extending the survival of T-effector cells.1 GITR is involved in the regulation of T-cell receptor mediated cell death. Constitutive expression of a transfected GITR gene induces resistance to apoptosis induced by anti CD3 monoclonal antibodies. GITR ligand interferes with the suppressor activity of CD4(+) CD25(+) regulatory T-cells.2
NCBI Gene Bank ID
References & Citations
1. Shimizu, J. et al. (2002) Nature Immunology 3:135–142.
2. Ji, Hb. et al. (2004)Journal of Immunology 172: 5823
Products are for research use only. Not for use in diagnostic or therapeutic procedures.