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MLH1 Recombinant Rabbit Monoclonal Antibody [PD01-61]

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Specification

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Catalog# HA721325

MLH1 Recombinant Rabbit Monoclonal Antibody [PD01-61]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

Overview

Product Name

MLH1 Recombinant Rabbit Monoclonal Antibody [PD01-61]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide corresponding to Human MLH1 aa 700-756.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P

Target Molecular Weight

Predicted band size: 85 kDa

Positive Control

HeLa cell lysate, HCT 116 cell lysate, A549 cell lysate, C2C12 cell lysate, NIH/3T3 cell lysate, human appendix tissue, human colon tissue, human B-cell lymphoblastic lymphoma tissue, human NKT cell lymphoma tissue, mouse testis tissue, mouse thymus tissue, human colon carcinoma tissue, rat small intestine tissue.

Conjugation

unconjugated

Clone Number

PD01-61

RRID

AB_3072442

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

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

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IHC-P

  • 1:200-1:1,000

Target

Function

DNA-mismatch repair (MMR) is an essential process in maintaining genetic stability. Lack of a functional DNA-mismatch repair pathway is a common characteristic of several different types of human cancers, either due to an MMR gene mutation or promoter methylation gene silencing. MLH1 is an integral part of the protein complex responsible for mismatch repair and is expressed in lymphocytes, heart, colon, breast, lung, spleen, testis, prostate, thyroid and gall bladder tissues, and is methylated in several ovarian tumors. Loss of MLH1 protein expression is associated with a mutated phenotype, microsatellite instability and a predisposition to cancer. In hereditary nonpolyposis colorectal cancer (HNPCC), an autosomal dominant inherited cancer syndrome that signifies a high risk of colorectal and various other types of cancer, the MLH1 gene exhibits a pathogenic mutation. Certain cancer cell lines, including leukemia CCRF-CEM, colon HCT 116 and KM12, and ovarian cancers SK-OV-3 and IGROV-1, show complete deficiency of MLH1, while MLH1 is expressed in 60% of melanomas, 70% of noninvasive squamous cell carcinomas and 30% of invasive squamous cell carcinomas.

Background References

1. Guan J et al. MLH1 Deficiency-Triggered DNA Hyperexcision by Exonuclease 1 Activates the cGAS-STING Pathway. Cancer Cell. 2021 Jan

2. Cannavo E et al. Regulation of the MLH1-MLH3 endonuclease in meiosis. Nature. 2020 Oct

Sequence Similarity

Belongs to the DNA mismatch repair MutL/HexB family.

Tissue Specificity

Colon, lymphocytes, breast, lung, spleen, testis, prostate, thyroid, gall bladder and heart.

Subcellular Location

Nucleus.

UNIPROT

SwissProt: P40692 Human

SwissProt: Q9JK91 Mouse

SwissProt: P97679 Rat

Synonyms

COCA 2 antibody

COCA2 antibody

DNA mismatch repair protein Mlh1 antibody

FCC 2 antibody

FCC2 antibody

hMLH 1 antibody

hMLH1 antibody

HNPCC 2 antibody

HNPCC antibody

HNPCC2 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of MLH1 on different lysates with Rabbit anti-MLH1 antibody (<a href="/products/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HCT 116 cell lysate (negative)<br />Lane 3: A549 cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 85 kDa<br />Observed band size: 85 kDa<br /><br />Exposure time: 2 minutes; ECL: K1801;<br /><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/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</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.

    ☑ Relative expression (RE)

    Western blot analysis of MLH1 on different lysates with Rabbit anti-MLH1 antibody (HA721325) at 1/2,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HCT 116 cell lysate (negative)
    Lane 3: A549 cell lysate

    Lysates/proteins at 15 µg/Lane.

    Predicted band size: 85 kDa
    Observed band size: 85 kDa

    Exposure time: 2 minutes; 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 (HA721325) 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.

  • Western blot analysis of MLH1 on different lysates with Rabbit anti-MLH1 antibody (<a href="/products/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</a>) at 1/2,000 dilution.<br /><br />Lane 1: C2C12 cell lysate<br />Lane 2: NIH/3T3 cell lysate<br /><br />Lysates/proteins at 30 µg/Lane.<br /><br />Predicted band size: 85 kDa<br />Observed band size: 85 kDa<br /><br />Exposure time: 1 minute; ECL: K1801;<br /><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/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</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 MLH1 on different lysates with Rabbit anti-MLH1 antibody (HA721325) at 1/2,000 dilution.

    Lane 1: C2C12 cell lysate
    Lane 2: NIH/3T3 cell lysate

    Lysates/proteins at 30 µg/Lane.

    Predicted band size: 85 kDa
    Observed band size: 85 kDa

    Exposure time: 1 minute; 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 (HA721325) 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.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of MLH1 on different lysates with Rabbit anti-MLH1 antibody (<a href="/products/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</a>) at 1/2,000 dilution.<br /><br />Lane 1: Hela-si NT cell lysate (10 µg/Lane)<br />Lane 2: Hela-si MLH1 cell lysate (10 µg/Lane)<br /><br />Predicted band size: 85 kDa<br />Observed band size: 85 kDa<br /><br />Exposure time: 3 minutes;<br />4-20% SDS-PAGE gel.<br /><br /><a href="/products/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</a> was shown to specifically react with MLH1 in Hela-si NT cells. Weakened band was observed when Hela-si MLH1 sample was tested. Hela-si NT and Hela-si MLH1 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</a>, 1/2,000) and Loading control antibody (Rabbit anti-GAPDH, <a href="/products/ET1601-4" style="font-weight: bold;text-decoration: underline;">ET1601-4</a>, 1/10,000) were 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.

    ☑ Knockdown (KD)

    Western blot analysis of MLH1 on different lysates with Rabbit anti-MLH1 antibody (HA721325) at 1/2,000 dilution.

    Lane 1: Hela-si NT cell lysate (10 µg/Lane)
    Lane 2: Hela-si MLH1 cell lysate (10 µg/Lane)

    Predicted band size: 85 kDa
    Observed band size: 85 kDa

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

    HA721325 was shown to specifically react with MLH1 in Hela-si NT cells. Weakened band was observed when Hela-si MLH1 sample was tested. Hela-si NT and Hela-si MLH1 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (HA721325, 1/2,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were 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 appendix tissue with Rabbit anti-MLH1 antibody (<a href="/products/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</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 appendix tissue with Rabbit anti-MLH1 antibody (HA721325) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721325) 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 colon tissue with Rabbit anti-MLH1 antibody (<a href="/products/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</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 colon tissue with Rabbit anti-MLH1 antibody (HA721325) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721325) 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 B-cell lymphoblastic lymphoma tissue with Rabbit anti-MLH1 antibody (<a href="/products/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</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 human B-cell lymphoblastic lymphoma tissue with Rabbit anti-MLH1 antibody (HA721325) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721325) 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 human NKT cell lymphoma tissue with Rabbit anti-MLH1 antibody (<a href="/products/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</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 NKT cell lymphoma tissue with Rabbit anti-MLH1 antibody (HA721325) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721325) 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 testis tissue with Rabbit anti-MLH1 antibody (<a href="/products/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</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 testis tissue with Rabbit anti-MLH1 antibody (HA721325) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721325) 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 thymus tissue with Rabbit anti-MLH1 antibody (<a href="/products/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</a>) at 1/200 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</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 mouse thymus tissue with Rabbit anti-MLH1 antibody (HA721325) at 1/200 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721325) 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 human colon carcinoma tissue with Rabbit anti-MLH1 antibody (<a href="/products/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</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 colon carcinoma tissue with Rabbit anti-MLH1 antibody (HA721325) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721325) 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 small intestine tissue with Rabbit anti-MLH1 antibody (<a href="/products/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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/HA721325" style="font-weight: bold;text-decoration: underline;">HA721325</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 rat small intestine tissue with Rabbit anti-MLH1 antibody (HA721325) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA721325) 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 colon cancer tissue using anti-MLH1 antibody (1/100) performed on the Dako Omnis.

    Immunohistochemical analysis of paraffin embedded human colon cancer tissue using anti-MLH1 antibody (1/100) performed on the Dako Omnis.

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