Recombinant
RabMAb

Recombinant Anti-MLH1 antibody [EPR20741] - BSA and Azide free (ab230383)

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Overview

  • Product name

    Anti-MLH1 antibody [EPR20741] - BSA and Azide free
    See all MLH1 primary antibodies

  • Description

    Rabbit monoclonal [EPR20741] to MLH1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, ICC/IF, Flow Cytmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment within Human MLH1 aa 350-800. The exact sequence is proprietary.
    Database link: P40692

  • Positive control

    • IHC-P: Human colon, colon cancer and breast cancer tissues. ICC/IF: SW480 cells. Flow: SW480 cells.

  • General notes

    Ab230383 is the carrier-free version of ab229191. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab230383 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab230383 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
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

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-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 specificity

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

  • Involvement in disease

    Defects 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 similarities

    Belongs to the DNA mismatch repair mutL/hexB family.

  • Cellular localization

    Nucleus.
  • 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

    Images

    • Flow Cytometry - Anti-MLH1 antibody [EPR20741] - BSA and Azide free (ab230383)
      Flow Cytometry - Anti-MLH1 antibody [EPR20741] - BSA and Azide free (ab230383)

      Flow cytometric analysis of HCT 116 (human colorectal carcinoma epithelial cell line, left) / SW480 (human colorectal adenocarcinoma cell line) cells labelling MLH1 with ab229191 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at 1/2000 dilution.

      Gated on total viable cells.

      The HCT 116 (human colorectal carcinoma epithelial) cell line is a negative control for MLH1 (PMID: 23724141)(left panel).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229191).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR20741] - BSA and Azide free (ab230383)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR20741] - BSA and Azide free (ab230383)

      Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labelling MLH1 with ab229191 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining in tumor cells of human breast cancer (PMID: 20215533) is observed. Counterstained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229191).

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR20741] - BSA and Azide free (ab230383)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR20741] - BSA and Azide free (ab230383)

      Immunohistochemical analysis of paraffin-embedded human colon tissue labelling MLH1 with ab229191 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining in human colon tissue is observed (PMID: 10535979). Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229191).

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-MLH1 antibody [EPR20741] - BSA and Azide free (ab230383)
      Immunocytochemistry/ Immunofluorescence - Anti-MLH1 antibody [EPR20741] - BSA and Azide free (ab230383)

      Immunofluorecent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SW480 (human colorectal adenocarcinoma cell line) cells labelling MLH1 with ab229191 at 1/500 dilution, followed by AlexaFluor®488 Goat anti-Rabbit (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in SW480 cell line. 

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

      Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889).

      The HCT 116 (human colorectal carcinoma epithelial) cell line is a negative control for MLH1 (PMID: 23724141).

      Secondary antibody only control: Used PBS instead of ab229191, followed by AlexaFluor®488 Goat anti-Rabbit secondary antibody (ab150077) at 1/1000 dilution.

       

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229191).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR20741] - BSA and Azide free (ab230383)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MLH1 antibody [EPR20741] - BSA and Azide free (ab230383)

      Immnohistochemical analysis of paraffin-embedded human colon cancer tissue labelling MLH1 with ab229191 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining in tumor cells of human colon cancer is observed (PMID: 22608206). Counter stained with hematoxylin.

      Secondary antibody only control: used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab229191).

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    References

    ab230383 has not yet been referenced specifically in any publications.

    Publishing research using ab230383? Please let us know so that we can cite the reference in this datasheet.

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