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TIM 3 Mouse Monoclonal Antibody [1E3]

Usd: 185

Specification

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Catalog# EM1701-19

TIM 3 Mouse Monoclonal Antibody [1E3]

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

Reactivity

  • Human

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

TIM 3 Mouse Monoclonal Antibody [1E3]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Recombinant protein with Human TIM 3 1-89 / 301.

Species Reactivity

Human

Validated Applications

WB, IHC-P, IF-Cell, FC

Molecular Weight

Predicted band size: 33 kDa

Positive Control

A549 cell lysate, HepG2 cell lysate, HDLM-2 cell lysate, human colon cancer tissue, human tonsil tissue, 293T, Daudi.

Conjugation

unconjugated

Clone Number

1E3

RRID

AB_3068747

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IHC-P

  • 1:1,000-1:5,000

  • FC

  • 1:50-1:100

Target

Function

Cell surface receptor implicated in modulating innate and adaptive immune responses. Generally accepted to have an inhibiting function. Reports on stimulating functions suggest that the activity may be influenced by the cellular context and/or the respective ligand. Regulates macrophage activation. Inhibits T-helper type 1 lymphocyte (Th1)-mediated auto- and alloimmune responses and promotes immunological tolerance. In CD8+ cells attenuates TCR-induced signaling, specifically by blocking NF-kappaB and NFAT promoter activities resulting in the loss of IL-2 secretion. The function may implicate its association with LCK proposed to impair phosphorylation of TCR subunits, and/or LGALS9-dependent recruitment of PTPRC to the immunological synapse.

Background References

1. Markwick LJ et al. Blockade of PD1 and TIM3 restores innate and adaptive immunity in patients with acute alcoholic hepatitis. Gastroenterology 148(3):590-602 (2015).

2. Delpoux A et al. FOXO1 opposition of CD8+ T cell effector programming confers early memory properties and phenotypic diversity. Proc Natl Acad Sci U S A 114(42):E8865-E8874 (2017).

Sequence Similarity

Belongs to the immunoglobulin superfamily. TIM family.

Tissue Specificity

Expressed in T-helper type 1 (Th1) lymphocytes. Expressed on regulatory T (Treg) cells after TCR stimulation. Expressed in dendritic cells and natural killer (NK) cells. Expressed in epithelial tissues. Expression is increased on CD4+ and CD8+ T-cells in chronic hepatitis C virus (HCV) infection. In progressive HIV-1 infection, expression is up-regulated on HIV-1-specific CD8 T-cells.

Post-translational Modification

O-glycosylated with core 1 or possibly core 8 glycans.; Phosphorylated on tyrosine residues; modestly increased after TCR/CD28 stimulation. Can be phosphorylated in the cytoplasmatic domain by FYN (By similarity). Phosphorylation at Tyr-265 is increased by stimulation with ligand LGALS9.

Subcellular Location

Membrane. Cell junction.

UNIPROT

SwissProt: Q8TDQ0 Human

Synonyms

CD366 antibody

FLJ14428 antibody

HAVcr-2 antibody

Havcr2 antibody

HAVR2_HUMAN antibody

Hepatitis A virus cellular receptor 2 antibody

Kidney injury molecule 3 antibody

KIM 3 antibody

KIM3 antibody

T cell immunoglobulin and mucin domain containing 3 antibody

Expand

Images

  • Western blot analysis of TIM 3 on different lysates with Mouse anti-TIM 3 antibody (<a href="/products/EM1701-19" style="font-weight: bold;text-decoration: underline;">EM1701-19</a>) at 1/2,000 dilution.<br /><br />Lane 1: A549 cell lysate (no heat)<br />Lane 2: HepG2 cell lysate (no heat)<br />Lane 3: HDLM-2 cell lysate<br /><br />Notice: no heat means the lysate is not boiled.<br /><br />Lysates/proteins at 30 µg/Lane.<br /><br />Predicted band size: 33 kDa<br />Observed band size: 42 kDa<br /><br />Exposure time: 3 minutes 10 seconds;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/EM1701-19" style="font-weight: bold;text-decoration: underline;">EM1701-19</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of TIM 3 on different lysates with Mouse anti-TIM 3 antibody (EM1701-19) at 1/2,000 dilution.

    Lane 1: A549 cell lysate (no heat)
    Lane 2: HepG2 cell lysate (no heat)
    Lane 3: HDLM-2 cell lysate

    Notice: no heat means the lysate is not boiled.

    Lysates/proteins at 30 µg/Lane.

    Predicted band size: 33 kDa
    Observed band size: 42 kDa

    Exposure time: 3 minutes 10 seconds;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1701-19) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Mouse anti-TIM 3 antibody (<a href="/products/EM1701-19" style="font-weight: bold;text-decoration: underline;">EM1701-19</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1701-19" style="font-weight: bold;text-decoration: underline;">EM1701-19</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Mouse anti-TIM 3 antibody (EM1701-19) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1701-19) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Mouse anti-TIM 3 antibody (<a href="/products/EM1701-19" style="font-weight: bold;text-decoration: underline;">EM1701-19</a>) at 1/5,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM1701-19" style="font-weight: bold;text-decoration: underline;">EM1701-19</a>) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Mouse anti-TIM 3 antibody (EM1701-19) at 1/5,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM1701-19) at 1/5,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunocytochemistry analysis of 293T cells labeling Tim3 with Mouse anti-Tim3 antibody (<a href="/products/EM1701-19" style="font-weight: bold;text-decoration: underline;">EM1701-19</a>) at 1/50 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-Tim3 antibody (<a href="/products/EM1701-19" style="font-weight: bold;text-decoration: underline;">EM1701-19</a>) at 1/50 dilution in 2% BSA overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

    Immunocytochemistry analysis of 293T cells labeling Tim3 with Mouse anti-Tim3 antibody (EM1701-19) at 1/50 dilution.

    Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-Tim3 antibody (EM1701-19) at 1/50 dilution in 2% BSA overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation