Clone:
REA1004
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MC
Alternative names:
VSIG9, Vstm3, WUCAM

Extended validation for TIGIT Antibody, anti-human, REAfinity™

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 REA1004
741182++
A15153G++
MBSA43++
Cells were incubated with an excess of purified unconjugated TIGIT (REA1004) 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 TIGIT. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with TIGIT antibodies and with a suitable counterstaining. As a control, TIGIT 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 TIGIT. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with TIGIT antibodies and with a suitable counterstaining. As a control, TIGIT 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 TIGIT. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with TIGIT antibodies and with a suitable counterstaining. As a control, TIGIT 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 TIGIT. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with TIGIT antibodies and with a suitable counterstaining. As a control, TIGIT 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 TIGIT. Human peripheral blood mononuclear cells (PBMCs) stimulated with CD3/CD28 antibodies (T Cell TransAct™) for 3 days were stained with TIGIT antibodies and with a suitable counterstaining. As a control, TIGIT 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 TIGIT (REA1004). 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 TIGIT (REA1004). 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 TIGIT (REA1004). 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 TIGIT Antibody, anti-human, REAfinity™

Overview

Clone REA1004 recognizes the human T cell immunoglobulin and ITIM domain (TIGIT) antigen, a protein containing an immunoglobulin variable (IgV) domain, a transmembrane domain, and an immunoreceptor tyrosine-based inhibitory motif (ITIM). TIGIT is expressed on T cells, including Treg cells and memory subsets, as well as on natural killer (NK) cells. The poliovirus receptor (PVR; also called NECL5 or CD155) is a high-affinity coreceptor for TIGIT that is highly expressed on dendritic cells (DCs), fibroblasts, endothelial cells, and some tumor cells. The interaction of TIGIT with PVR on DCs leads to the down-regulation of pro-inflammatory cytokine secretion and the induction of interleukin 10 (IL-10) production in DCs, exerting an inhibitory effect on T cell activation.
Additional information: Clone REA1004 displays negligible binding to Fc receptors.

Alternative names

VSIG9, Vstm3, WUCAM

Detailed product information

Technical specifications

CloneREA1004
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenTIGIT
Alternative names of antigenVSIG9, Vstm3, WUCAM
Molecular mass of antigen [kDa]24
Distribution of antigendendritic cells, NK cells, T cells, tumor cells, memory T cells, regulatory T cells
Entrez Gene ID201633
RRIDAB_2751336, AB_2751348, AB_2751337, AB_2751349, AB_2751338, AB_2751350, AB_2751339, AB_2751351, AB_2751340, AB_2751352, AB_2751341, AB_2801891, AB_2751347

Resources for TIGIT Antibody, anti-human, REAfinity™

Documents and Protocols

References for TIGIT Antibody, anti-human, REAfinity™

Publications

  1. Yu, X. et al. (2009) The surface protein TIGIT suppresses T cell activation by promoting the generation of mature immunoregulatory dendritic cells. Nat. Immunol. 10(1): 48-57
  2. Levin, S. D. et al. (2011) Vstm3 is a member of the CD28 family and an important modulator of T-cell function. Eur. J. Immunol. 41(4): 902-915

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