Clone:
DT5D3
Type of antibody:
Primary antibodies, Functional-grade antibodies
Isotype:
mouse IgG1κ, mouse IgG1
Applications:
FC, MICS, IF, IHC, FA
Alternative names:
TNFRSF18, AITR, GITR-D

Extended validation for CD357 (GITR) Antibody, anti-human

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with DT5D3
110416++
108-17++
AITR++
REA1007++
Cells were incubated with an excess of purified unconjugated CD357 (GITR) (DT5D3) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for CD357 (GITR). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD357 (GITR) antibodies and with a suitable counterstaining. As a control, CD357 (GITR) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD357 (GITR). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD357 (GITR) antibodies and with a suitable counterstaining. As a control, CD357 (GITR) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD357 (GITR). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD357 (GITR) antibodies and with a suitable counterstaining. As a control, CD357 (GITR) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD357 (GITR). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD357 (GITR) antibodies and with a suitable counterstaining. As a control, CD357 (GITR) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for CD357 (GITR). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD357 (GITR) antibodies and with a suitable counterstaining. As a control, CD357 (GITR) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for CD357 (GITR). Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with CD357 (GITR) antibodies and with a suitable counterstaining. As a control, CD357 (GITR) antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, and cell doublets were excluded from the analysis based on scatter signals and 4',6-diamidino-2-phenylindole (DAPI) fluorescence. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using CD357 (GITR) (DT5D3). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using CD357 (GITR) (DT5D3). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using CD357 (GITR) (DT5D3). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for CD357 (GITR) Antibody, anti-human

Overview

CD357, also known as glucocorticoid-induced tumor necrosis factor receptor–related protein (GITR) or TNFRSF18, is an inducible type I transmembrane protein and member of the tumor necrosis factor receptor (TNFR) superfamily.
CD357 is expressed at low levels on thymocyte subsets, resting T cells, B cells, macrophages, and at high levels on CD4
+
CD25
+
regulatory T cells (Tregs). Upon activation, expression on CD4
+
and CD8
+
T cells is upregulated. Triggering of CD357 has been described to modulate Treg function and costimulate effector T cells. Stimulation of T cells through CD357 can abrogate the inhibitory function of Tregs. It is hypothesized that CD357 has a role in the maintenance of immunological self tolerance, and mouse models of autoimmune disease suggest that CD357 activation may break self-tolerance and induce autoimmunity.

Alternative names

TNFRSF18, AITR, GITR-D

Detailed product information

Technical specifications

CloneDT5D3
Clonalitymonoclonal
Isotypemouse IgG1κ, mouse IgG1
Isotype controlIsotype Control Antibody, mouse IgG1
Hostmouse
Type of antibodyPrimary antibodies, Functional-grade antibodies
Specieshuman
AntigenCD357 (GITR)
Alternative names of antigenTNFRSF18, AITR, GITR-D
Molecular mass of antigen [kDa]24
Entrez Gene ID8784
RRIDAB_2751597, AB_2784153, AB_2784152, AB_2801819, AB_2801818, AB_1036234, AB_871549, AB_2751643

References for CD357 (GITR) Antibody, anti-human

Publications

  1. Shimizu, J. et al. (2002)
    Stimulation of CD25
    +
    CD4
    +
    regulatory T cells through GITR breaks immunological self-tolerance.
    Nat. Immunol. 3(2): 135-142
  2. Nocentini, G. et al. (1997) 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. 94: 6216-6221
  3. Nocentini, G. et al. (2007) GITR/GITRL: more than an effector T cell co-stimulatory system. Eur. J. Immunol. 37: 1165-1169
  4. Gurney, A. L. et al. (1999) Identification of a new member of the tumor necrosis factor family and its receptor, a human ortholog of mouse GITR. Curr. Biol. 9: 215-218

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