Overview

  • Product nameAnti-MSH6 antibody [EPR3945]
    See all MSH6 primary antibodies
  • Description
    Rabbit monoclonal [EPR3945] to MSH6
  • Tested applicationsWB, IHC-P, IHC, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human MSH6 aa 1-100 (N terminal).

  • Positive control
    • WB: A431, HeLa and SW480 cell lysates IHC-P: Human colonic adenocarcinoma tissue ICC/IF: HeLa cells
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels.

    If you have any questions regarding this update, please contact our Scientific Support team.

     

    Alternative versions available:

    Anti-MSH6 antibody (Alexa Fluor® 647) [EPR3945] (ab198334)

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Dissociation constant (KD) KD = 2.30 x 10 -9 M Learn more about KD
  • Storage bufferpH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • PurityProtein A purified
  • Purification notesThis antibody is not purified. It is provided in cell supernatant and storage buffer.
  • Clonality Monoclonal
  • Clone numberEPR3945
  • IsotypeIgG
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab92471 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 1/1000 - 1/10000. Predicted molecular weight: 163 kDa.
IHC-P 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

For unpurified, use 1/100 - 1/250.

IHC Use at an assay dependent concentration. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF 1/250.

Target

  • FunctionComponent of the post-replicative DNA mismatch repair system (MMR). Heterodimerizes with MSH2 to form MutS alpha, which binds to DNA mismatches thereby initiating DNA repair. When bound, MutS alpha bends the DNA helix and shields approximately 20 base pairs, and recognizes single base mismatches and dinucleotide insertion-deletion loops (IDL) in the DNA. After mismatch binding, forms a ternary complex with the MutL alpha heterodimer, which is thought to be responsible for directing the downstream MMR events, including strand discrimination, excision, and resynthesis. ATP binding and hydrolysis play a pivotal role in mismatch repair functions. The ATPase activity associated with MutS alpha regulates binding similar to a molecular switch: mismatched DNA provokes ADP-->ATP exchange, resulting in a discernible conformational transition that converts MutS alpha into a sliding clamp capable of hydrolysis-independent diffusion along the DNA backbone. This transition is crucial for mismatch repair. MutS alpha may also play a role in DNA homologous recombination repair.
  • Involvement in diseaseDefects in MSH6 are the cause of hereditary non-polyposis colorectal cancer type 5 (HNPCC5) [MIM:600678]. 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. 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. MSH6 mutations appear to be associated with atypical HNPCC and in particular with development of endometrial carcinoma or atypical endometrial hyperplasia, the presumed precursor of endometrial cancer. Defects in MSH6 are also found in familial colorectal cancers (suspected or incomplete HNPCC) that do not fulfill the Amsterdam criteria for HNPCC.
    Defects in MSH6 are a cause of susceptibility to endometrial cancer (ENDMC) [MIM:608089].
  • Sequence similaritiesBelongs to the DNA mismatch repair mutS family.
    Contains 1 PWWP domain.
  • Post-translational
    modifications
    The N-terminus is blocked.
    Phosphorylated upon DNA damage, probably by ATM or ATR.
    Phosphorylated by PRKCZ, which may prevent MutS alpha degradation by the ubiquitin-proteasome pathway.
  • Cellular localizationNucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • DNA mismatch repair protein MSH6 antibody
    • G/T mismatch binding protein antibody
    • G/T mismatch-binding protein antibody
    • GTBP antibody
    • GTMBP antibody
    • hMSH6 antibody
    • HNPCC 5 antibody
    • HNPCC5 antibody
    • HSAP antibody
    • MSH 6 antibody
    • MSH6 antibody
    • MSH6_HUMAN antibody
    • mutS (E. coli) homolog 6 antibody
    • MutS alpha 160 kDa subunit antibody
    • MutS homolog 6 (E. coli) antibody
    • mutS homolog 6 antibody
    • MutS-alpha 160 kDa subunit antibody
    • p160 antibody
    • Sperm associated protein antibody
    see all

Anti-MSH6 antibody [EPR3945] images

  • Immunohistochemical staining of paraffin embedded rat liver with purified ab92471 at a dilution of 1/500. A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunohistochemical staining of paraffin embedded rat liver with unpurified ab92471 at a dilution of 1/150. A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunohistochemical staining of paraffin embedded human colon with unpurified ab92471 at a dilution of 1/500. A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunohistochemical staining of paraffin embedded human colon with unpurified ab92471 at a dilution of 1/150. A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunofluorescent staining of HeLa cells (fixed in 4% PFA, permeabilized with 0.1% Triton X 100) using purified ab92471 at a dilution of 1/250. An Alexa Fluor® 488 goat anti-rabbit antibody was used as the secondary at a dilution of 1/500 and the cells were counter stained with DAPI. The negative control is shown in the bottom right hand panel - for the negative control, Alex Fluor® 488 goat anti-mouse was used at a dilution of 1/500.

  • Anti-MSH6 antibody [EPR3945] (ab92471) at 1/1000 dilution (purified) + Rat brain at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size : 163 kDa
    Observed band size : 160 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • purified at 1/6000 dilution + SW480 cell lysate at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size : 163 kDa
    Observed band size : 160 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Anti-MSH6 antibody [EPR3945] (ab92471) at 1/2000 dilution (unpurified) + SW480 cell lysate at 10 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size : 163 kDa
    Observed band size : 160 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Unpurified ab92471, at a 1/100 dilution, detecting MSH6 in paraffin embedded Human colonic adenocarcinoma tissue by immunohistochemistry. Detection used HRP conjugated anti rabbit antibody.

  • Unpurified ab92471 (1/500) staining MSH6 in asynchronous 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 see abreview.

    See Abreview

  • All lanes : Anti-MSH6 antibody [EPR3945] (ab92471) at 1/1000 dilution (unpurified)

    Lane 1 : A431 cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : SW480 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP labelled goat anti-rabbit antibody at 1/2000 dilution

    Predicted band size : 163 kDa

    Secondary antibody - goat anti-rabbit HRP (ab6721)

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References for Anti-MSH6 antibody [EPR3945] (ab92471)

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) »
  • Tsai JH  et al. Traditional serrated adenoma has two pathways of neoplastic progression that are distinct from the sessile serrated pathway of colorectal carcinogenesis. Mod Pathol N/A:N/A (2014). Read more (PubMed: 24603588) »

See all 8 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)
Antigen retrieval step Heat mediated
Sample Human Tissue sections (endometrial cancer)
Specification endometrial cancer
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 23 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Lymphoblastoid cell line)
Loading amount 20 µg
Specification Lymphoblastoid cell line
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted May 15 2013

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 12 2012

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