Validated using a knockout cell line

Anti-MSH6 antibody [44] (ab14204)

Reviews (2)Q&A (2)References (2)

Key features and details

  • Mouse monoclonal [44] to MSH6
  • Suitable for: ICC/IF, WB, IP, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Dog, Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-MSH6 antibody [44]
    See all MSH6 primary antibodies

  • Description

    Mouse monoclonal [44] to MSH6

  • Host species

    Mouse

  • Tested applications

    Suitable for: ICC/IF, WB, IP, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Dog, Human

  • Immunogen

    Synthetic peptide, corresponding to amino acids 225-333 of Human MSH6

  • Positive control

    • WB: HeLa, A431 and HAP1 (HAP1-MSH6 knockout cell lysate used as negative control) cell lysates. IHC-P: Human colon carcinoma tissue. ICC/IF: HeLa cells and HAP1 cells (HAP1-MSH6 knockout cells used as negative cell line).

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.

  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituent: 1% BSA
  • Concentration information loading...

  • Purity

    Protein G purified

  • Clonality

    Monoclonal

  • Clone number

    44

  • Isotype

    IgG1

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab14204 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
ICC/IF 1/250.
WB 1/100 - 1/500.
IP 1/100 - 1/500.
IHC-P Use at an assay dependent concentration.

Target

  • Function

    Component 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 disease

    Defects 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 similarities

    Belongs 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 localization

    Nucleus.
  • Information by UniProt

  • Database links

    see all

  • 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

Images

  • Western blot - Anti-MSH6 antibody [44] (ab14204)
    Western blot - Anti-MSH6 antibody [44] (ab14204)
    All lanes : Anti-MSH6 antibody [44] (ab14204)

    Lane 1 : Hap1 cell lysate
    Lane 2 : HEK-293 cell lysate
    Lane 3 : Wild-type HeLa cell lysate
    Lane 4 : MSH6 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.


    Lanes 1 - 4: Merged signal (red and green). Green - ab14204 observed at 160 kDa. Red - loading control, ab52866 observed at 50 kDa.

     ab14204 was shown to react with MSH6 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255410 (knockout cell lysate ab263763) was used. Wild-type and MSH6 knockout samples were subjected to SDS-PAGE. ab14204 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

  • Western blot - Anti-MSH6 antibody [44] (ab14204)
    Western blot - Anti-MSH6 antibody [44] (ab14204)
    All lanes : Anti-AMACR + p63 antibody [4A4 (p63)] (ab14202)

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : MSH6 knockout HAP1 cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : A431 cell lysate

    Lysates/proteins at 20 µg per lane.


    Lanes 1 - 4: Merged signal (red and green). Green - ab14204 observed at 160 kDa. Red - loading control, ab18251, observed at 52 kDa.


    ab14204 was shown to specifically react with MSH6 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when MSH6 knockout HAP1 samples were used. Wild-type and MSH6 knockout samples were subjected to SDS-PAGE. ab14204 and ab18251 (loading control to alpha Tubulin) were diluted at 1/100 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [44] (ab14204)
    Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [44] (ab14204)

    ab14204 staining MSH6 in wild-type HAP1 cells (top panel) and MSH6 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab14204 at 1/250 dilution and ab202272 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [44] (ab14204)
    Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [44] (ab14204)

    ab14204 staining MSH6 in HeLa cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab14204 at 1/250dilution and ab202272 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [44] (ab14204)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [44] (ab14204)
    ab14204 staining human colon carcinoma by IHC-P.

References

Publishing research using ab14204? Please let us know so that we can cite the reference in this datasheet.

ab14204 has been referenced in 2 publications.

  • McKinney JA  et al. Distinct DNA repair pathways cause genomic instability at alternative DNA structures. Nat Commun 11:236 (2020). PubMed: 31932649
  • Steigerwald C  et al. Sensitization of colorectal cancer cells to irinotecan by the Survivin inhibitor LLP3 depends on XAF1 proficiency in the context of mutated p53. Arch Toxicol 92:2645-2648 (2018). PubMed: 29947891

Customer reviews and Q&As

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1-4 of 4 Abreviews or Q&A

Question

Please send me certificate of analysis for the following items,


MLH-1 Lot *********– AB16878



MSH-6 Lot ********1 – AB14204


Thank you,

Read More
Answer

Thank you for contacting Abcam.
Please find attached the Certificate of Compliances for ab14204 and ab16878 as requested.
If there is anything else I can help you with, please let me know.

Read More

Western blot abreview for Anti-MSH6 antibody [44]

 Average
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse Embryonic Fibroblast)
Loading amount
25 µg
Specification
Mouse Embryonic Fibroblast
Gel Running Conditions
Reduced Denaturing (10% SDS PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Feb 11 2009

Western blot abreview for Anti-MSH6 antibody [44]

 Good
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Loading amount
25 µg
Specification
HeLa
Gel Running Conditions
Reduced Denaturing (10% SDS PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

Submitted Feb 11 2009

Question

I thought that the antigen was a peptide (amino acids 225-333), based on information for clone 44 from [a competitor]. Could you confirm that Abcam used full length human Msh6, and explain the discrepancy? Thank you

Read More
Answer

Thank you for your email. I apologize as there was an error on the online datasheet for this antibody. The immunogen used was in fact a synthetic peptide, corresponding to amino acids 225-333 of Human MSH6. This has been updated, and thank you for bringing this to our attention. Please contact us again if you have any additional questions.

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