Anti-GITR (Glucocorticoid-induced TNFR-related Protein, GITR-D, Activation-inducible TNFR Family Rec

Glucocorticoid induced Tumor Necrosis Factor receptor family related gene (GITR, AITR) is a type 1 228 amino acid transmembrane protein that has been designated TNFRSF18. GITR is expressed mainly by CD4+ CD25+ T lymphocytes. Its engagement by antibodies or its ligand (GITRL) induces a costimulatory signal which can break T cell suppression (2). GITR has been implicated as a regulatory mechanism in parasitic immune resposes (4), and is involved with Macrophages activation in atherosclerotic plaque formation (3). Applications: Suitable for use in FLISA and Flow Cytometry. Other applications not tested. Recommended Dilution: Flow Cytometry: 1:50, 1.6ul of concentrated G2035-08W labels 5x10e5 cells, Optimal dilutions to be determined by the researcher. Flow Cytometry: Ficoll prepared human peripheral blood leukocytes (PBL) were stimulated 5 days in culture with PHA. , 1. 5x10e5 cells per tube were washed and pre-incubated 5 minutes with 20ul of 250ug/ml human IgG (to block non specific binding), after which they were incubated 45 minutes on ice with 80ul of G2035-08W at a 1:50 dilution (10ug/ml). 2. Cells were washed three times and analyzed by FACS. Cells stained positive with a mean shift of 1.3 log10 fluorescent units when compared to a mouse IgG3-R-PE negative control at a similar concentration. , 3. Binding was blocked when cells were pre-incubated with a 10-fold excess of G2035-08S (unlabeled anti-GITR mAb). Storage and Stability: Store product at 4°C in the dark. DO NOT FREEZE! Stable at 4°C for 12 months after receipt as an undiluted liquid. Dilute required amount only prior to immediate use. Further dilutions can be made in assay buffer. Caution: PE conjugates are sensitive to light. For maximum recovery of product, centrifuge the original vial prior to removing the cap.