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Flow cytometric analysis of GITR (CD357) expression on human peripheral blood mononuclear cells. Human peripheral blood mononuclear cells were cultured for 3 days in medium with Phytohemagglutinin-M (PHA-M Stimulated; 1.5% v/v). The cells were then stained with either Purified Mouse IgG2b, κ Isotype Control (Cat. No. 555740; dashed line histogram) or Purified Mouse Anti-Human GITR (CD357) antibody (Cat. No. 567769; solid line histogram) followed by PE Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 550589). The fluorescent histogram showing GITR (CD357) expression (or Ig Isotype staining) was derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometry and data analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System and FloJo™ software.
BD Pharmingen™ Purified Mouse Anti-Human GITR (CD357)
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
The V27-580 monoclonal antibody specifically binds to GITR (Glucocorticoid-Induced Tumor necrosis factor Receptor), a member of the tumor necrosis factor receptor (TNFR) superfamily that is designated TNFRSF18. In the human, GITR is expressed at low levels in peripheral blood T cells, bone marrow, thymus, spleen, and lymph nodes and is up-regulated upon antigen stimulation or by treatment with anti-CD3 plus anti-CD28. GITR is also reported to be constitutively expressed on Treg cells. GITR's ligand (GITRL) is a member of the TNF superfamily, is designated TNFSF18, and is expressed on antigen presenting cells. The GITR cytoplasmic domain has striking homology with the cytoplasmic domains of the co-stimulatory receptors CD137 (4-1BB), CD134 (OX40) and CD27. GITR signaling is mediated by signaling adaptors, TNFR-associated factors (TRAFs), that affect signaling pathways (eg, Erk, JNK, MAPK and NF-κB) to enhance T-cell survival and cytokine production. The effects of GITR signaling upon the dynamic and interconnected roles of effector and regulatory T lymphocytes in the immune response are under investigation.
Development References (7)
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Clouthier DL, Watts TH. Cell-specific and context-dependent effects of GITR in cancer, autoimmunity, and infection.. Cytokine Growth Factor Rev. 2014; 25(2):91-106. (Biology). View Reference
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Kwon B, Yu KY, Ni J, et al. Identification of a novel activation-inducible protein of the tumor necrosis factor receptor superfamily and its ligand.. J Biol Chem. 1999; 274(10):6056-61. (Biology). View Reference
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Li Z, Mahesh SP, Kim BJ, Buggage RR, Nussenblatt RB. Expression of glucocorticoid induced TNF receptor family related protein (GITR) on peripheral T cells from normal human donors and patients with non-infectious uveitis.. J Autoimmun. 2003; 21(1):83-92. (Biology). View Reference
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Nocentini G, Giunchi L, Ronchetti S, et al. A new member of the tumor necrosis factor / nerve growth factor receptor family inhibits T cell receptor-induced apoptosis... Proc Natl Acad Sci U S A. 1997; 94(12):6216-6221. (Biology: Northern blot). View Reference
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Pedroza-Gonzalez A, Zhou G, Singh SP, et al. GITR engagement in combination with CTLA-4 blockade completely abrogates immunosuppression mediated by human liver tumor-derived regulatory T cells ex vivo.. Oncoimmunology. 2015; 4(12):e1051297. (Biology: Flow cytometry). View Reference
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Placke T, Kopp HG, Salih HR. Glucocorticoid-induced TNFR-related (GITR) protein and its ligand in antitumor immunity: functional role and therapeutic modulation.. Clin Dev Immunol. 2010; 2010:239083. (Biology). View Reference
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Shevach EM, Stephens GL. The GITR-GITRL interaction: co-stimulation or contrasuppression of regulatory activity?. Nat Rev Immunol. 2006; 6(8):613-8. (Biology). View Reference
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