Anti tim3 clone rmt3 23

Manufactured by BioLegend

Anti-TIM3 (clone RMT3-23) is a monoclonal antibody that recognizes the T cell immunoglobulin and mucin domain-containing protein 3 (TIM-3). TIM-3 is an inhibitory receptor expressed on various immune cells and is involved in the regulation of immune responses.

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RMT3-23 (7 citations)

2 protocols using anti tim3 clone rmt3 23

Evaluating Immune Checkpoint Expression

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Mouse spleen cells were washed with PBS containing 1% BSA (staining buffer), blocked with mouse IgG in staining buffer, then stained with conjugated antibodies. Antibodies used were anti-PD-1 (clone 29F.1A12, Biolegend, San Diego, CA), anti-TIGIT (clone 1G9, Biolegend), anti-GITR (clone yGITR765, Biolegend), anti-CD25 (clone PC61, Biolegend), anti-PD-L1 (clone 10F.9G2, Biolegend), anti-CTLA4 (clone UC10-4B9, Biolegend), anti-2B4 (clone M2B4(B6)458.1, Biolegend), anti-VISTA (clone MIH63, Biolegend), anti-LAG3 (clone C9B7W, Biolegend), anti-TIM3 (clone RMT3-23, Biolegend), and anti-CD4 (clone RM4-5, Biolegend).
Human PBMCs were stained with anti-CD4 (clone OKT4, Biolegend), anti-CD25 (clone BC96, Biolegend), anti-FoxP3 (clone 236A/E7, Biolegend), anti-PD-1 (clone EH12.2H7, Biolegend), anti-TIGIT (clone A15153G, Biolegend). Prior to anti-FoxP3 staining, the cells were fixed for 18 hours and permeabilized.

Muhammad F., Wang D., McDonald T., Walsh M., Drenen K., Montieth A., Foster C.S, & Lee D.J. (2020). TIGIT+ A2Ar-Dependent Anti-Uveitic Treg Cells are a Novel Subset of Tregs Associated with Resolution of Autoimmune Uveitis. Journal of autoimmunity, 111, 102441.

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Comprehensive Splenic T Cell Analysis

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Spleens were harvested after mice were sacrificed 48 hours after CLP, and then were processed to single-cell suspensions though a 70 μm filter. Splenocytes were rinsed with 10 mL cold PBS, and 200 μL from each spleen was put into a 96-well plate for staining. Anti-CD3 (BD, clone 500A2), anti-CD4 (clone RM4-5, BioLegend), anti-CD8 (clone MCD0830, Invitrogen), anti-CD44 ( clone IM7, BioLegend), anti-CD226 (clone 10E5, BioLegend), anti-PD-1 (clone 29F.1A12, BioLegend), anti-2B4 (clone eBio244F4, Thermo Fisher Scientific), and anti-TIM-3 (clone RMT3-23, BioLegend) were used for surface staining to determine T cell phenotype. For the detection of cell apoptosis, splenocytes were stained with a FITC Annexin V apoptosis detection kit with 7-AAD (BioLegend). Tregs were identified via intracellular staining for Foxp3-APC (clone FJK-16S, eBioscience). Splenocytes were surface stained for anti-CD62L (MEL-14), anti-CD69 (H1.2F3), and then permeabilized using a Foxp3 kit (BD Biosciences) and stained with anti-Foxp3, anti-CTLA-4 (UC10-489), and anti-Helios (22F6, all Abs from BioLegend). AccuCheck Counting Beads (Thermo Fisher Scientific) were added after staining to calculate the absolute number of cells per spleen.

Sun Y., Anyalebechi J.C., Sun H., Yumoto T., Xue M., Liu D., Liang Z., Coopersmith C.M, & Ford M.L. (2021). Anti-TIGIT differentially affects sepsis survival in immunologically experienced versus previously naive hosts. JCI Insight, 6(5), e141245.

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