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FITC Rat Anti-Mouse CD357 (GITR)
FITC Rat Anti-Mouse CD357 (GITR)

Flow cytometric analysis of CD357 (GITR) expression on mouse CD4-positive splenocytes. C57BL/6 splenocytes were stained with APC Rat anti-Mouse CD25 (Cat. No. 557192), PE Rat anti-Mouse CD4 (Cat. No. 553048/553049), and either FITC Rat IgG2b Isotype Control (Cat. No. 553988; open histograms) or FITC Rat anti-Mouse CD357 (GITR) (Cat. No. 558139; shaded histograms).  Viable leukocytes were selected by exclusion of propidium iodide (Cat. No. 556463).  The left panel represents the expression of GITR by CD25-positive/CD4-positive T lymphocytes (Treg cells), and the right  panel shows CD25-negative/CD4-positive T lymphocytes.  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Flow cytometric analysis of CD357 (GITR) expression on mouse CD4-positive splenocytes. C57BL/6 splenocytes were stained with APC Rat anti-Mouse CD25 (Cat. No. 557192), PE Rat anti-Mouse CD4 (Cat. No. 553048/553049), and either FITC Rat IgG2b Isotype Control (Cat. No. 553988; open histograms) or FITC Rat anti-Mouse CD357 (GITR) (Cat. No. 558139; shaded histograms).  Viable leukocytes were selected by exclusion of propidium iodide (Cat. No. 556463).  The left panel represents the expression of GITR by CD25-positive/CD4-positive T lymphocytes (Treg cells), and the right  panel shows CD25-negative/CD4-positive T lymphocytes.  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.

Product Details
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BD Pharmingen™
GITR; Gitr; glucocorticoid-induced TNFR-related protein; Tnfrsf18; AITR
Mouse (QC Testing)
Rat IgG2b
Mouse CD25+ CD4+ T Cell Line
Flow cytometry (Routinely Tested)
0.5 mg/ml
21936
AB_647178
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with FITC under optimum conditions, and unreacted FITC was removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
558139 Rev. 3
Antibody Details
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DTA-1

The DTA-1 monoclonal antibody specifically binds to GITR [Glucocorticoid-induced Tumor necrosis factor (TNF) receptor family-Related], a 66-70-kDa homodimer glycoprotein that is a member of the TNF receptor superfamily and is also known as TNFRSF18 and CD357. As its name implies, GITR expression was first detected in T lymphocytes that had been treated with dexamethasone, a glucocorticoid. In normal naive mice, GITR is expressed at moderate levels on CD25-positive/CD4-positive/CD8a-negative thymocytes and on CD25-positive/CD4-positive/CD45RB-low splenocytes. It is also expressed at low levels on splenic CD25-negative/CD4-positive/CD45RB-low T lymphocytes, B lymphocytes, macrophages, and dendritic cells. Activation of T and B lymphocytes upregulates GITR expression. GITR is a costimulatory receptor that plays an important role in Regulatory T (Treg)-cell functions, and a GITR Ligand has been detected on B lymphocytes, macrophages, and dendritic cells. mAb DTA-1 abrogates suppression by Treg cells without affecting their proliferative response, while it is co-stimulatory for T lymphocytes that are not Treg cells.

558139 Rev. 3
Format Details
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FITC
Fluorescein (FITC) is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 518-nm. FITC is designed to be excited by the Blue laser (488-nm) and detected using an optical filter centered near 520 nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
FITC
Blue 488 nm
494 nm
518 nm
558139 Rev.3
Citations & References
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Development References (10)

  1. Beilharz MW, Sammels LM, Paun A, et al. Timed ablation of regulatory CD4-positive T cells can prevent murine AIDS progression. J Immunol. 2004; 172:4917-4925. (Clone-specific). View Reference
  2. Dittmer U, He H, Messer RJ, et al. Functional impairment of CD8-positive T cells by regulatory T cells during persistent retroviral infection. Immunity. 2004; 20:293-303. (Clone-specific). View Reference
  3. Ji H, Liao G, Faubion WA, et al. The natural ligand for glucocorticoid-induced TNF receptor-related protein abrogates regulatory T cell suppression. J Immunol. 2004; 172:5823-5827. (Biology). View Reference
  4. Ko K, Yamazaki S, Nakamura K, et al. Treatment of advanced tumors with agonistic anti-GITR mAb and its effects on tumor-infiltrating Foxp3+CD25+CD4+ regulatory T cells. J Exp Med. 2005; 202(7):885-891. (Clone-specific). View Reference
  5. Kohm AP, Williams JS, Miller SD. Ligation of the glucocorticoid-induced TNF receptor enhances autoreactive CD4-positive T cell activation and experimental autoimmune encephalomyelitis. J Immunol. 2004; 172:4686-4690. (Clone-specific). View Reference
  6. 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; 94:6216-6221. (Biology). View Reference
  7. Shimizu J, Moriizumi E. CD4-positive CD25-negative T cells in aged mice are hyporesponsive and exihibit suppressive activity. J Immunol. 2003; 170:1675-1682. (Clone-specific). View Reference
  8. Shimizu J, Yamazaki S, Takahashi T, Ishida Y, Sakaguchi S. Stimulation of CD25-positive CD4-positive regulatory T cells through GITR breaks immunological self-tolerance. Nat Immunol. 2002; 3(2):135-142. (Immunogen). View Reference
  9. Tone M, Tone Y, Adams E, et al. Mouse glucocorticoid-induced tumor necrosis factor receptor ligand is costimulatory for T cells. Proc Natl Acad Sci U S A. 2003; 100(25):15059-15064. (Biology). View Reference
  10. Uraushihara K, Kanai T, Ko K, et al. Regulation of murine inflammatory bowel disease by CD25-positive and CD25negative CD4-positive glucocorticoid-induced TNF receptor family-related gene-positive regulatory T cells. J Immunol. 2003; 171:708-716. (Clone-specific). View Reference
View All (10) View Less
558139 Rev. 3

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