Clone:
RMT3-23
Type of antibody:
Primary antibodies
Isotype:
rat IgG2aκ
Applications:
FC, MICS, IF, IHC
Alternative names:
HAVCR2, TIMD-3, Tim3

Extended validation for TIM-3 Antibody, anti-mouse

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 RMT3-23
REA602++
5D12-
215015 (rIgG2a)-
215008R (rIgG2a)++
Cells were incubated with an excess of purified unconjugated TIM-3 (RMT3-23) 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 TIM-3. Splenocytes from CD57BL/6 mice were stained with TIM-3 antibodies and with a suitable counterstaining. As a control, TIM-3antibody 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 TIM-3. Splenocytes from CD57BL/6 mice were stained with TIM-3 antibodies and with a suitable counterstaining. As a control, TIM-3antibody 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 TIM-3. Splenocytes from CD57BL/6 mice were stained with TIM-3 antibodies and with a suitable counterstaining. As a control, TIM-3antibody 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 TIM-3. Splenocytes from CD57BL/6 mice were stained with TIM-3 antibodies and with a suitable counterstaining. As a control, TIM-3antibody 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 TIM-3. Splenocytes from CD57BL/6 mice were stained with TIM-3 antibodies and with a suitable counterstaining. As a control, TIM-3antibody 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 TIM-3. Splenocytes from CD57BL/6 mice were stained with TIM-3 antibodies and with a suitable counterstaining. As a control, TIM-3antibody 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 TIM-3 (RMT3-23). 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 TIM-3 (RMT3-23). 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 TIM-3 (RMT3-23). 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 TIM-3 Antibody, anti-mouse

Overview

The monoclonal antibody RMT3-23 reacts with mouse TIM-3, which is a Th-1-specific cell surface protein expressed by differentiated CD4
+
Th1 and CD8
+
Tc1 cells and macrophages. It is absent on CD4
+
Th2 and CD8
+
Tc2 cells. TIM-3 has been proposed to inhibit Th1-mediated immune responses. TIM-3 absence enhances the severity of experimental autoimmune encephalomyelitis in mice. The interaction of TIM-3 and Galectin-9 induces apoptosis of Th1 cells.

Alternative names

HAVCR2, TIMD-3, Tim3

Detailed product information

Technical specifications

CloneRMT3-23
Clonalitymonoclonal
Isotyperat IgG2aκ
Isotype controlIsotype Control Antibody, rat IgG2a
Hostrat
Type of antibodyPrimary antibodies
Speciesmouse
AntigenTIM-3
Alternative names of antigenHAVCR2, TIMD-3, Tim3
Molecular mass of antigen [kDa]29
Entrez Gene ID171285
RRIDAB_2654180, AB_2654181, AB_2654179

Resources for TIM-3 Antibody, anti-mouse

Certificates

Please follow this
link
to search for Certificates of Analysis (CoA) by lot number.

References for TIM-3 Antibody, anti-mouse

Publications

  1. Monney, L. et al. (2002) Tʜ1-specific cell surface protein Tim-3 regulates macrophage activation and severity of an autoimmune disease. Nature 415: 536-541
  2. van de Weyer, P. S. et al. (2006) A highly conserved tyrosine of Tim-3 is phosphorylated upon stimulation by its ligand galectin-9. Biochem. Biophys. Res. Commun. 351(2): 571-576

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