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TIM 3 Recombinant Antibody [PSH13-95] - Mouse IgG1 (Chimeric)

Usd: 350

Specification

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Catalog# HA601442

TIM 3 Recombinant Antibody [PSH13-95] - Mouse IgG1 (Chimeric)

Application

  • WB

  • IF-Cell

  • FC

Reactivity

  • Human

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

TIM 3 Recombinant Antibody [PSH13-95] - Mouse IgG1 (Chimeric)

Antibody Type

Recombinant Chimeric Antibody

Immunogen

Recombinant protein within human TIM 3 aa 1-223.

Species Reactivity

Human

Validated Applications

WB, IF-Cell, FC

Molecular Weight

Predicted band size: 33 kDa

Positive Control

RPMI 8226 cell lysate, RPMI 8226.

Conjugation

unconjugated

Clone Number

PSH13-95

Reactivity Data

WB IF-Cell FC
Human
Tested Tested
Tested Tested
Tested Tested

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

Target

Function

Hepatitis A virus cellular receptor 2 (HAVCR2), also known as T-cell immunoglobulin and mucin-domain containing-3 (TIM-3), is a protein that in humans is encoded by the HAVCR2 (TIM-3)gene. HAVCR2 was first described in 2002 as a cell surface molecule expressed on IFNγ producing CD4+ Th1 and CD8+ Tc1 cells. Later, the expression was detected in Th17 cells, regulatory T-cells, and innate immune cells (dendritic cells, NK cells, monocytes, macrophages). HAVCR2 receptor is a regulator of the immune response.

Background References

1. Kandel S et al. The TIM3/Gal9 signaling pathway: An emerging target for cancer immunotherapy. Cancer Lett. 2021 Jul

2. Palacios LM et al. TIM3 Expression in Anaplastic-Thyroid-Cancer-Infiltrating Macrophages: An Emerging Immunotherapeutic Target. Biology (Basel). 2022 Nov

Subcellular Location

Membrane, Cell junction, Cell membrane.

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

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of TIM 3 on different lysates with Mouse anti-TIM 3 antibody (<a href="/products/HA601442" style="font-weight: bold;text-decoration: underline;">HA601442</a>) at 1/2,000 dilution.<br /><br />Lane 1: RPMI 8226 cell lysate (20 µg/Lane)<br />Lane 2: HT-29 cell lysate (negative) (20 µg/Lane)<br /><br />Predicted band size: 33 kDa<br />Observed band size: 55 kDa<br />Exposure time: 59 seconds; ECL: K1802;<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/HA601442" style="font-weight: bold;text-decoration: underline;">HA601442</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. 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.

    ☑ Relative expression (RE)

    Western blot analysis of TIM 3 on different lysates with Mouse anti-TIM 3 antibody (HA601442) at 1/2,000 dilution.

    Lane 1: RPMI 8226 cell lysate (20 µg/Lane)
    Lane 2: HT-29 cell lysate (negative) (20 µg/Lane)

    Predicted band size: 33 kDa
    Observed band size: 55 kDa
    Exposure time: 59 seconds; ECL: K1802;
    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 (HA601442) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Immunocytochemistry analysis of RPMI 8226 (positive) and HT-29 (negative) labeling TIM 3 with Mouse anti-TIM 3 antibody (<a href="/products/HA601442" style="font-weight: bold;text-decoration: underline;">HA601442</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-TIM 3 antibody (<a href="/products/HA601442" style="font-weight: bold;text-decoration: underline;">HA601442</a>) at 1/100 dilution in 1% BSA in PBST 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. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) were used as the secondary antibody at 1/1,000 dilution.

    ☑ Relative expression (RE)

    Immunocytochemistry analysis of RPMI 8226 (positive) and HT-29 (negative) labeling TIM 3 with Mouse anti-TIM 3 antibody (HA601442) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-TIM 3 antibody (HA601442) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Flow cytometric analysis of HT-29 (left, negative) and RPMI 8226 (right, positive) cells labeling TIM 3.<br /><br />Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (<a href="/products/HA601442" style="font-weight: bold;text-decoration: underline;">HA601442</a>, 1/1,000) (red) compared with Mouse IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Mouse IgG Secondary antibody (<a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    ☑ Relative expression (RE)

    Flow cytometric analysis of HT-29 (left, negative) and RPMI 8226 (right, positive) cells labeling TIM 3.

    Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA601442, 1/1,000) (red) compared with Mouse IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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