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TIM 3 Recombinant Rabbit Monoclonal Antibody [PSH16-50]

Usd: 385

Specification

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Catalog# HA723854

TIM 3 Recombinant Rabbit Monoclonal Antibody [PSH16-50]

Application

  • WB

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

TIM 3 Recombinant Rabbit Monoclonal Antibody [PSH16-50]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IP

Molecular Weight

Predicted band size: 33 kDa

Positive Control

RPMI 8226 cell lysate, RAW264.7 cell lysates, human lung cancer tissue, mouse spleen tissue, rat spleen tissue.

Conjugation

unconjugated

Clone Number

PSH16-50

Reactivity Data

WB IHC-P IP
Human
Tested Tested
Tested Tested
Tested Tested
Mouse
Tested Tested
Tested Tested
Rat
Not Recommended
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.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:200-1:1,000

  • IP

  • 1-2μg/sample

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

SwissProt: Q8VIM0 Mouse

SwissProt: P0C0K5 Rat

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 Rabbit anti-TIM 3 antibody (<a href="/products/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</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 /><br />Exposure time: 5 seconds; ECL: K1801;<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/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</a>) at 1/2,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</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 Rabbit anti-TIM 3 antibody (HA723854) 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: 5 seconds; ECL: K1801;
    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 (HA723854) at 1/2,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of TIM 3 on RAW264.7 cell lysates with Rabbit anti-TIM 3 antibody (<a href="/products/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</a>) at 1/2,000 dilution.<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 33 kDa<br />Observed band size: 70 kDa<br />Exposure time: 1 minute; 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/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of TIM 3 on RAW264.7 cell lysates with Rabbit anti-TIM 3 antibody (HA723854) at 1/2,000 dilution.

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 33 kDa
    Observed band size: 70 kDa
    Exposure time: 1 minute; 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 (HA723854) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Rabbit anti-TIM 3 antibody (<a href="/products/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</a>) at 1/1,000 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/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</a>) at 1/1,000 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 lung cancer tissue with Rabbit anti-TIM 3 antibody (HA723854) at 1/1,000 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 (HA723854) at 1/1,000 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 mouse spleen tissue with Rabbit anti-TIM 3 antibody (<a href="/products/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</a>) at 1/1,000 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/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</a>) at 1/1,000 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 mouse spleen tissue with Rabbit anti-TIM 3 antibody (HA723854) at 1/1,000 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 (HA723854) at 1/1,000 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 rat spleen tissue with Rabbit anti-TIM 3 antibody (<a href="/products/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</a>) at 1/200 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/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</a>) at 1/200 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 rat spleen tissue with Rabbit anti-TIM 3 antibody (HA723854) at 1/200 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 (HA723854) at 1/200 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.

  • TIM 3 was immunoprecipitated from 0.2 mg RPMI 8226 cell lysate with <a href="/products/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</a> at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: RPMI 8226 cell lysate (input)<br />Lane 2: <a href="/products/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</a> IP in RPMI 8226 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</a> in RPMI 8226 cell lysate<br /><br />Blocking/Dilution buffer: primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>)<br />Exposure time: 3 seconds; ECL: <a href="/products/K1801" style="font-weight: bold;text-decoration: underline;">K1801</a>

    TIM 3 was immunoprecipitated from 0.2 mg RPMI 8226 cell lysate with HA723854 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723854 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: RPMI 8226 cell lysate (input)
    Lane 2: HA723854 IP in RPMI 8226 cell lysate
    Lane 3: Rabbit IgG instead of HA723854 in RPMI 8226 cell lysate

    Blocking/Dilution buffer: primary antibody dilution (K1803)
    Exposure time: 3 seconds; ECL: K1801

  • TIM 3 was immunoprecipitated from 0.2 mg RAW264.7 cell lysate with <a href="/products/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</a> at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: RAW264.7 cell lysate (input)<br />Lane 2: <a href="/products/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</a> IP in RAW264.7 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA723854" style="font-weight: bold;text-decoration: underline;">HA723854</a> in RAW264.7 cell lysate<br /><br />Blocking/Dilution buffer: primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>)<br />Exposure time: 46 seconds; ECL: <a href="/products/K1802" style="font-weight: bold;text-decoration: underline;">K1802</a>

    TIM 3 was immunoprecipitated from 0.2 mg RAW264.7 cell lysate with HA723854 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723854 at 1/2,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: RAW264.7 cell lysate (input)
    Lane 2: HA723854 IP in RAW264.7 cell lysate
    Lane 3: Rabbit IgG instead of HA723854 in RAW264.7 cell lysate

    Blocking/Dilution buffer: primary antibody dilution (K1803)
    Exposure time: 46 seconds; ECL: K1802

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