Overview

  • Product nameAnti-MLH1 antibody [EPR3894]See all MLH1 primary antibodies ...
  • Description
    Rabbit monoclonal [EPR3894] to MLH1
  • Tested applicationsWB, IP, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    A synthetic peptide corresponding to residues in Human MLH1 (UniProt ID: P40692).

  • Positive control
    • 293T, HeLa, A431, and SW480 cell lysates; Human colonic adenocarcinoma and tonsil tissues.
  • General notesProduced under U.S. Patent No. 5,675,063.

Properties

Applications

Our Abpromise guarantee covers the use of ab92312 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/10000 - 1/50000. Predicted molecular weight: 84 kDa.
IP 1/10 - 1/100.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. PubMed: 22022465Performed after boiling twice before IHC protocol carried out.
Flow Cyt 1/10 - 1/100.
ICC/IF 1/200.

Target

  • FunctionHeterodimerizes 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-MSH6) 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.
  • Tissue specificityColon, lymphocytes, breast, lung, spleen, testis, prostate, thyroid, gall bladder and heart.
  • Involvement in diseaseDefects in MLH1 are the cause of hereditary non-polyposis colorectal cancer type 2 (HNPCC2) [MIM:609310]. Mutations in more than one gene locus can be involved alone or in combination in the production of the HNPCC phenotype (also called Lynch syndrome). Most families with clinically recognized HNPCC have mutations in either MLH1 or MSH2 genes. HNPCC is an autosomal, dominantly inherited disease associated with marked increase in cancer susceptibility. It is characterized by a familial predisposition to early onset colorectal carcinoma (CRC) and extra-colonic cancers of the gastrointestinal, urological and female reproductive tracts. HNPCC is reported to be the most common form of inherited colorectal cancer in the Western world, and accounts for 15% of all colon cancers. Cancers in HNPCC originate within benign neoplastic polyps termed adenomas. Clinically, HNPCC is often divided into two subgroups. Type I: hereditary predisposition to colorectal cancer, a young age of onset, and carcinoma observed in the proximal colon. Type II: patients have an increased risk for cancers in certain tissues such as the uterus, ovary, breast, stomach, small intestine, skin, and larynx in addition to the colon. Diagnosis of classical HNPCC is based on the Amsterdam criteria: 3 or more relatives affected by colorectal cancer, one a first degree relative of the other two; 2 or more generation affected; 1 or more colorectal cancers presenting before 50 years of age; exclusion of hereditary polyposis syndromes. The term 'suspected HNPCC' or 'incomplete HNPCC' can be used to describe families who do not or only partially fulfill the Amsterdam criteria, but in whom a genetic basis for colon cancer is strongly suspected.
    Defects in MLH1 are a cause of mismatch repair cancer syndrome (MMRCS) [MIM:276300]; also known as Turcot syndrome or brain tumor-polyposis syndrome 1 (BTPS1). MMRCS is an autosomal dominant disorder characterized by malignant tumors of the brain associated with multiple colorectal adenomas. Skin features include sebaceous cysts, hyperpigmented and cafe au lait spots.
    Defects in MLH1 are a cause of Muir-Torre syndrome (MuToS) [MIM:158320]; also abbreviated MTS. MuToS is a rare autosomal dominant disorder characterized by sebaceous neoplasms and visceral malignancy.
    Note=Defects in MLH1 may contribute to lobular carcinoma in situ (LCIS), a non-invasive neoplastic disease of the breast.
    Defects in MLH1 are a cause of susceptibility to endometrial cancer (ENDMC) [MIM:608089].
    Note=Some epigenetic changes can be transmitted unchanged through the germline (termed 'epigenetic inheritance'). Evidence that this mechanism occurs in humans is provided by the identification of individuals in whom 1 allele of the MLH1 gene is epigenetically silenced throughout the soma (implying a germline event). These individuals are affected by HNPCC but does not have identifiable mutations in MLH1, even though it is silenced, which demonstrates that an epimutation can phenocopy a genetic disease.
  • Sequence similaritiesBelongs to the DNA mismatch repair mutL/hexB family.
  • Cellular localizationNucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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
    • MGC5172 antibody
    • MLH 1 antibody
    • MLH1 antibody
    • MLH1_HUMAN antibody
    • MutL homolog 1 (E. coli) antibody
    • MutL homolog 1 antibody
    • MutL homolog 1 colon cancer nonpolyposis type 2 antibody
    • MutL homolog 1, colon cancer, nonpolyposis type 2 (E. coli) antibody
    • MutL protein homolog 1 antibody
    • MutL, E. coli, homolog of, 1 antibody
    see all

Anti-MLH1 antibody [EPR3894] images

  • All lanes : Anti-MLH1 antibody [EPR3894] (ab92312) at 1/10000 dilution

    Lane 1 : 293T cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : A431 cell lysate
    Lane 4 : SW480 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    goat anti-rabbit HRP at 1/2000 dilution

    Predicted band size : 84 kDa
  • ab92312 at 1/100 dilution staining MLH1 in Human colonic adenocarcinoma by Immunohistochemistry, Paraffin-embedded tissue. The use of an HRP/AP polymerized antibody is recommended for a secondary antibody.
  • ab92312 (1/200) staining MLH1 in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to abreview.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab92312 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab92312, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • ab92312 staining MLH1 in Human colorectal (top) and gastric tissue (bottom) by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
  • ab92312 at 1/100 dilution staining MLH1 in Human tonsil by Immunohistochemistry, Paraffin-embedded tissue. The use of an HRP/AP polymerized antibody is recommended for a secondary antibody.

References for Anti-MLH1 antibody [EPR3894] (ab92312)

This product has been referenced in:
  • Morales C  et al. Immune chaperone gp96 drives the contributions of macrophages to inflammatory colon tumorigenesis. Cancer Res 74:446-59 (2014). Mouse . Read more (PubMed: 24322981) »
  • Gerdes MJ  et al. Highly multiplexed single-cell analysis of formalin-fixed, paraffin-embedded cancer tissue. Proc Natl Acad Sci U S A 110:11982-7 (2013). ICC/IF ; Human . Read more (PubMed: 23818604) »

See all 4 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Antigen retrieval step Heat mediated
Sample Human Tissue sections (endometrial cancer)
Specification endometrial cancer
Permeabilization No
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 23 2013

Merci de confirmer ces détails et de votre coopération. Les détails fournis nous permettent de suivre de près la qualité de nos produits.

Je suis désolé que ces produits ne fonctionnent pas co...

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Merci beaucoup pour toutes ces informations détaillées qui nous sont très utiles pour identifier l'origine des problèmes rencontrés avec ab92312 et ab126057.

Je n'ai rien à modifier à votre prot...

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Je suis désolée d'apprendre que les anticorps ab92312 et ab126057 ne vous donnent pas de bons résultats en WB.

Pour nous permettre de vous fournir la meilleure assistance technique, pourriez-vous nous donner les informatio...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton-X100 in PBS
Username

Dr. Kirk McManus

Verified customer

Submitted Apr 18 2012

Thank you for contacting us.
I can confirm that the immunogen sequence of ab92312 is 100% similar with chicken MLH1 (http://www.uniprot.org/uniprot/E1BQE0)
ab62729; the sequence is 81% similar to chicken MLH1 (http://www.uniprot.org/uniprot/E...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Various tumor cell lines)
Loading amount 30 µg
Specification Various tumor cell lines
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Nov 03 2010

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