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MLH1 Mouse Monoclonal Antibody [A9F7]

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Catalog# HA601135

MLH1 Mouse Monoclonal Antibody [A9F7]

Application

  • WB

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

概述

产品名称

MLH1 Mouse Monoclonal Antibody [A9F7]

抗体类型

Mouse Monoclonal Antibody

免疫原

Recombinant protein within human MLH1 aa 452-751 / 751.

种属反应性

Human, Mouse, Rat

验证应用

WB, IHC-P

分子量

Predicted band size: 85 kDa

阳性对照

HeLa cell lysate, A549 cell lysate, HepG2 cell lysate, Jurkat cell lysate, NIH/3T3 cell lysate, F9 cell lysate, RAW264.7 cell lysate, C6 cell lysate, A431 cell lysate, Daudi cell lysate, K-562 cell lysate, HL-60 cell lysate, SW480 cell lysate, human colon carcinoma tissue, mouse colon tissue, rat small intestine tissue.

偶联

unconjugated

克隆号

A9F7

RRID

AB_3071857

产品特性

形态

Liquid

浓度

1ug/ul

存放说明

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

存储缓冲液

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

亚型

IgG2b

纯化方式

Protein A affinity purified.

应用稀释度

  • WB

  • 1:1,000-1:2,000

  • IHC-P

  • 1:1,000

靶点

功能

DNA mismatch repair protein Mlh1 or MutL protein homolog 1 is a protein that in humans is encoded by the MLH1 gene located on chromosome 3. It is a gene commonly associated with hereditary nonpolyposis colorectal cancer. Orthologs of human MLH1 have also been studied in other organisms including mouse and the budding yeast Saccharomyces cerevisiae. This gene was identified as a locus frequently mutated in hereditary nonpolyposis colon cancer. It is a human homolog of the E. coli DNA mismatch repair gene, mutL, which mediates protein-protein interactions during mismatch recognition, strand discrimination, and strand removal. Defects in MLH1 are associated with the microsatellite instability observed in hereditary nonpolyposis colon cancer. Alternatively spliced transcript variants encoding different isoforms have been described, but their full-length natures have not been determined.

背景文献

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

亚细胞定位

Nucleus, Chromosome.

UNIPROT #

SwissProt: P40692 Human

SwissProt: Q9JK91 Mouse

SwissProt: P97679 Rat

别名

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

展开

图片

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of MLH1 on different lysates with Mouse anti-MLH1 antibody (<a href="/products/HA601135" style="font-weight: bold;text-decoration: underline;">HA601135</a>) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.<br /><br />Lane 1: HeLa cell lysate (15 µg/Lane)<br />Lane 2: HCT 116 cell lysate (negative) (15 µg/Lane)<br />Lane 3: A549 cell lysate (15 µg/Lane)<br />Lane 4: HepG2 cell lysate (15 µg/Lane)<br />Lane 5: Jurkat cell lysate (15 µg/Lane)<br /><br />Predicted band size: 85 kDa<br />Observed band size: 85 kDa<br /><br />Exposure time: Lane 1-5 (left): 24 seconds; Lane 1-5 (right): 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/HA601135" style="font-weight: bold;text-decoration: underline;">HA601135</a>) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were 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 MLH1 on different lysates with Mouse anti-MLH1 antibody (HA601135) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.

    Lane 1: HeLa cell lysate (15 µg/Lane)
    Lane 2: HCT 116 cell lysate (negative) (15 µg/Lane)
    Lane 3: A549 cell lysate (15 µg/Lane)
    Lane 4: HepG2 cell lysate (15 µg/Lane)
    Lane 5: Jurkat cell lysate (15 µg/Lane)

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

    Exposure time: Lane 1-5 (left): 24 seconds; Lane 1-5 (right): 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 (HA601135) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were 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.

  • Western blot analysis of MLH1 on different lysates with Mouse anti-MLH1 antibody (<a href="/products/HA601135" style="font-weight: bold;text-decoration: underline;">HA601135</a>) at 1/1,000 dilution.<br /><br />Lane 1: NIH/3T3 cell lysate (20 µg/Lane)<br />Lane 2: F9 cell lysate (20 µg/Lane)<br />Lane 3: RAW264.7 cell lysate (20 µg/Lane)<br />Lane 4: C6 cell lysate (20 µg/Lane)<br /><br />Predicted band size: 85 kDa<br />Observed band size: 85 kDa<br /><br />Exposure time: 3 minutes 10 seconds; 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/HA601135" style="font-weight: bold;text-decoration: underline;">HA601135</a>) at 1/1,000 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 MLH1 on different lysates with Mouse anti-MLH1 antibody (HA601135) at 1/1,000 dilution.

    Lane 1: NIH/3T3 cell lysate (20 µg/Lane)
    Lane 2: F9 cell lysate (20 µg/Lane)
    Lane 3: RAW264.7 cell lysate (20 µg/Lane)
    Lane 4: C6 cell lysate (20 µg/Lane)

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

    Exposure time: 3 minutes 10 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 (HA601135) at 1/1,000 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.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of MLH1 on different lysates with Mouse anti-MLH1 antibody (<a href="/products/HA601135" style="font-weight: bold;text-decoration: underline;">HA601135</a>) at 1/1,000 dilution.<br /><br />Lane 1: A431 cell lysate (20 µg/Lane)<br />Lane 2: HeLa cell lysate (20 µg/Lane)<br />Lane 3: Daudi cell lysate (20 µg/Lane)<br />Lane 4: K-562 cell lysate (20 µg/Lane)<br />Lane 5: HL-60 cell lysate (20 µg/Lane)<br />Lane 6: SW480 cell lysate (20 µg/Lane)<br />Lane 7: HCT 116 cell lysate (Negative) (20 µg/Lane)<br /><br />Predicted band size: 85 kDa<br />Observed band size: 85 kDa<br /><br />Exposure time: 20 seconds;<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/HA601135" style="font-weight: bold;text-decoration: underline;">HA601135</a>) at 1/1,000 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 1:150,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of MLH1 on different lysates with Mouse anti-MLH1 antibody (HA601135) at 1/1,000 dilution.

    Lane 1: A431 cell lysate (20 µg/Lane)
    Lane 2: HeLa cell lysate (20 µg/Lane)
    Lane 3: Daudi cell lysate (20 µg/Lane)
    Lane 4: K-562 cell lysate (20 µg/Lane)
    Lane 5: HL-60 cell lysate (20 µg/Lane)
    Lane 6: SW480 cell lysate (20 µg/Lane)
    Lane 7: HCT 116 cell lysate (Negative) (20 µg/Lane)

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

    Exposure time: 20 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 (HA601135) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:150,000 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 Mouse anti-MLH1 antibody (<a href="/products/HA601135" style="font-weight: bold;text-decoration: underline;">HA601135</a>) at 1/2,000 dilution.<br /><br />Lane 1: Hela-si NT cell lysate<br />Lane 2: Hela-si MLH1 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 85 kDa<br />Observed band size: 85 kDa<br /><br />Exposure time: 50 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br /><a href="/products/HA601135" style="font-weight: bold;text-decoration: underline;">HA601135</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/HA601135" style="font-weight: bold;text-decoration: underline;">HA601135</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-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.

    ☑ Knockdown (KD)

    Western blot analysis of MLH1 on different lysates with Mouse anti-MLH1 antibody (HA601135) at 1/2,000 dilution.

    Lane 1: Hela-si NT cell lysate
    Lane 2: Hela-si MLH1 cell lysate

    Lysates/proteins at 10 µg/Lane.

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

    Exposure time: 50 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    HA601135 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 (HA601135, 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-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 carcinoma tissue with Mouse anti-MLH1 antibody (<a href="/products/HA601135" style="font-weight: bold;text-decoration: underline;">HA601135</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) 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/HA601135" style="font-weight: bold;text-decoration: underline;">HA601135</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 Mouse anti-MLH1 antibody (HA601135) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (HA601135) 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 colon tissue with Mouse anti-MLH1 antibody (<a href="/products/HA601135" style="font-weight: bold;text-decoration: underline;">HA601135</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) 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/HA601135" style="font-weight: bold;text-decoration: underline;">HA601135</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 colon tissue with Mouse anti-MLH1 antibody (HA601135) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (HA601135) 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 Mouse anti-MLH1 antibody (<a href="/products/HA601135" style="font-weight: bold;text-decoration: underline;">HA601135</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) 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/HA601135" style="font-weight: bold;text-decoration: underline;">HA601135</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 Mouse anti-MLH1 antibody (HA601135) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (HA601135) 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.

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