Overview

  • Product nameAnti-MLH1 antibody [G168-15]See all MLH1 primary antibodies ...
  • Description
    Mouse monoclonal [G168-15] to MLH1
  • Tested applicationsFlow Cyt, ICC/IF, IHC-P more details
  • Species reactivity
    Reacts with: Mouse, Rat, Hamster, Human
  • Immunogen

    Recombinant full length protein.

  • Positive control
    • Tonsil, colon carcinoma. This antibody gave a positive result in IF/ICC when used in the following formaldehyde fixed cell lines: HepG2

Properties

Applications

Our Abpromise guarantee covers the use of ab14206 in the following tested applications.

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

Application Notes
Flow Cyt 1/50.
ICC/IF 1/30. PubMed: 17543860
IHC-P 1/25 - 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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.
  • Target information above from: UniProt accession P40692 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    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 [G168-15] images

  • Formalin fixed paraffin embedded human tonsil stained with MLH1 using ABC and DAB chromogen.
  • Overlay histogram showing HeLa cells stained with ab14206 (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 (ab14206, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Immunohistochemical analysis of formalin-fixed, paraffin-embedded Human colon carcinoma tissue, staining MLH1 with ab14206.
  • ICC/IF image of ab14206 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab14206 at 1/50 dilution overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-MLH1 antibody [G168-15] (ab14206)

This product has been referenced in:
  • Bhattacharyya T  et al. Mechanistic basis of infertility of mouse intersubspecific hybrids. Proc Natl Acad Sci U S A 110:E468-77 (2013). Mouse . Read more (PubMed: 23329330) »
  • Borodin PM  et al. X-Y chromosome synapsis and recombination in 3 vole species of Asian lineage of the genus Microtus (Rodentia: Arvicolinae). Cytogenet Genome Res 132:129-33 (2011). ICC/IF . Read more (PubMed: 21042015) »

See all 4 Publications for this product

Product Wall

Merci de nous avoir contactés.

Je suis désolé d'apprendre que le produit ab14206 que vous avez reçu ne fonctionne pas comme attendu. Les informations que vous m'avez fournies vont nous permettre de chercher l'origin...

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Thank you for your call today and for letting us know about the trouble with this antibody.

As we discussed, I'm sending a free of charge vial of ab92312 on the order *** which should arrive shortly.

Please keep me updated about the...

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Thank you for contacting us.
The immunogen for ab14206 is the full length protein. However, the epitope of this antibody is not mapped. We do not know the sequence homology of the epitope with the chicken protein. The homology of the full length p...

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Thank you for your response. I have not had a chance to use MLH1 in my experiments. I recommend changes in fixation based on Abcams recommended protocols for ICC-IF. We recommend that users fix samples either in ice-cold methanol, acetone (1-10 min...

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Thank you for contacting Abcam regarding this issue. I am sorry to here of the problems that you have been experiencing using ab14206. Often, changing the fixation step of your tissue can lead to better results. A methanol fixation may work well for ...

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I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of a different lot of ab14206. To check the status of the order please c...

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Thanks for the additional information. I agree that the vial of ab14206 you received is not working as expected. If you have purchased this antibody within the last 6 months or so I would be happy to offer a replacement, credit or refund. In yo...

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Thank you for your enquiry. I am sorry to hear ab14206 is giving you trouble. MLH1 should be detected in human testis using this antibody so I am curious if you are experiencing high background, non-specific binding (or both) or just not seeing...

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Thank you for the images. Does your sentence "Besides the same second antibody can be work in another IHC detection with Cat#ab3366" mean that the secondary antibody works well? The colon tissue must be colon carcinoma, not normal colon tissue as th...

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Thank you for your enquiry. Yes, this antibody has been applied against rat cells using immunohistochemistry (ABC method). I am not aware of its application against spermatocytes, nor the expression profile of MLH1. However, I am assuming its expre...

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