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MLH1 Recombinant Antibody [PSH20-67] - Rabbit IgG (Chimeric)

Usd: 385

Specification

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Catalog# HA724342

MLH1 Recombinant Antibody [PSH20-67] - Rabbit IgG (Chimeric)

Application

  • IHC-P

  • IF-Cell

  • WB

Reactivity

  • Human

Conjugation

  • unconjugated

Overview

Product Name

MLH1 Recombinant Antibody [PSH20-67] - Rabbit IgG (Chimeric)

Antibody Type

Recombinant Chimeric Antibody

Species Reactivity

Human

Validated Applications

IHC-P, IF-Cell, WB

Target Molecular Weight

Predicted band size: 85 kDa

Positive Control

HeLa (Human cervical adenocarcinoma cells) cell lysate, SW480 (Human colorectal zdenocarcinoma cells) cell lysate.

Conjugation

unconjugated

Clone Number

PSH20-67

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

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

Purification Method

Protein A affinity purified.

Application Dilution

  • IHC-P

  • 1:2,000

  • IF-Cell

  • 1:3,000

  • WB

  • 1:2,000

Target

Function

Heterodimerizes with PMS2 to form MutL alpha, a component of the post-replicative DNA mismatch repair system (MMR). DNA repair is initiated by MutS alpha (MSH2-MSH6) or MutS beta (MSH2-MSH3) binding to a dsDNA mismatch, then MutL alpha is recruited to the heteroduplex. Assembly of the MutL-MutS-heteroduplex ternary complex in presence of RFC and PCNA is sufficient to activate endonuclease activity of PMS2. It introduces single-strand breaks near the mismatch and thus generates new entry points for the exonuclease EXO1 to degrade the strand containing the mismatch. DNA methylation would prevent cleavage and therefore assure that only the newly mutated DNA strand is going to be corrected. MutL alpha (MLH1-PMS2) interacts physically with the clamp loader subunits of DNA polymerase III, suggesting that it may play a role to recruit the DNA polymerase III to the site of the MMR. Also implicated in DNA damage signaling, a process which induces cell cycle arrest and can lead to apoptosis in case of major DNA damages. Heterodimerizes with MLH3 to form MutL gamma which plays a role in meiosis.

Background References

1. Yue Q et al. Histone H3K9 Lactylation Confers Temozolomide Resistance in Glioblastoma via LUC7L2-Mediated MLH1 Intron Retention. Adv Sci (Weinh). 2024 May

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

Subcellular Location

Nucleus, Chromosome.

UNIPROT

SwissProt: P40692 Human

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

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Human <br />Tissue: Tonsil <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0) (high pressure), 2 minutes. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA724342" style="font-weight: bold;text-decoration: underline;">HA724342</a>, 1/2,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Tonsil
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0) (high pressure), 2 minutes.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724342, 1/2,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P) <br /> <br />Species: Human <br />Tissue: Appendix <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0) (high pressure), 2 minutes. <br /> <br />Wash buffer: 1× TBST <br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature. <br />Primary antibody: <a href="/products/HA724342" style="font-weight: bold;text-decoration: underline;">HA724342</a>, 1/2,000, 1 hour at room temperature. <br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Appendix
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0) (high pressure), 2 minutes.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA724342, 1/2,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Application: Immunocytochemistry (IF-cell)<br /><br />Species: Human<br />Sample: SW480 (Human colorectal zdenocarcinoma cells)/HCT 116 (Human colon cancer cells)<br /><br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.<br />Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.<br /><br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.<br />Antibody dilution buffer: 1% BSA in PBST.<br />Primary antibody: <a href="/products/HA724342" style="font-weight: bold;text-decoration: underline;">HA724342</a>, 1/3,000, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 45 minutes at room temperature.<br /><br />Counterstain: Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).<br />Negative control: HCT 116 (PMID: 27618981).

    ☑ Relative expression (RE)

    Application: Immunocytochemistry (IF-cell)

    Species: Human
    Sample: SW480 (Human colorectal zdenocarcinoma cells)/HCT 116 (Human colon cancer cells)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.

    Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
    Antibody dilution buffer: 1% BSA in PBST.
    Primary antibody: HA724342, 1/3,000, overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.

    Counterstain: Beta tubulin (HA601187, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).
    Negative control: HCT 116 (PMID: 27618981).

  • <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/HA724342" style="font-weight: bold;text-decoration: underline;">HA724342</a>) at 1/2,000 dilution.<br /><br />Lane 1: HeLa (Human cervical adenocarcinoma cells) cell lysate<br />Lane 2: HCT 116 (Human colon cancer cells) cell lysate (negative)<br />Lane 3: SW480 (Human colorectal zdenocarcinoma cells) cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br />Exposure time: 12 seconds; ECL: K1801<br /><br />HCT 116 is a negative control (PMID: 34787019).<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/HA724342" style="font-weight: bold;text-decoration: underline;">HA724342</a>, 1/2,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 85 kDa<br />Observed band size: 85 kDa

    ☑ Relative expression (RE)

    Western blot analysis of MLH1 on different lysates with Rabbit anti-MLH1 antibody (HA724342) at 1/2,000 dilution.

    Lane 1: HeLa (Human cervical adenocarcinoma cells) cell lysate
    Lane 2: HCT 116 (Human colon cancer cells) cell lysate (negative)
    Lane 3: SW480 (Human colorectal zdenocarcinoma cells) cell lysate

    Lysates/proteins at 10 µg/Lane.
    Exposure time: 12 seconds; ECL: K1801

    HCT 116 is a negative control (PMID: 34787019).

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724342, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

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

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