Anti-MSH6 antibody [44] (ab14204)
- Datasheet
- References (1)
- Protocols
Overview
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Product nameAnti-MSH6 antibody [44]
See all MSH6 primary antibodies -
DescriptionMouse monoclonal [44] to MSH6
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Host speciesMouse
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Tested applicationsSuitable for: ICC/IF, WB, IP, IHC-Pmore details
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Species reactivityReacts with: Mouse, Rat, Dog, Human
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Immunogen
Synthetic peptide, corresponding to amino acids 225-333 of Human MSH6
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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
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.05% Sodium azide
Constituent: 1% BSA -
Concentration information loading... -
PurityProtein G purified
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ClonalityMonoclonal
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Clone number44
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IsotypeIgG1
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Research areas
Associated products
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Compatible Secondaries
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Isotype control
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
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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.
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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
modificationsThe 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
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Database links
- Entrez Gene: 474585 Dog
- Entrez Gene: 2956 Human
- Entrez Gene: 17688 Mouse
- Entrez Gene: 100360342 Rat
- Omim: 600678 Human
- SwissProt: P52701 Human
- SwissProt: P54276 Mouse
- Unigene: 445052 Human
see all -
Alternative names
- DNA mismatch repair protein Msh6 antibody
- G/T mismatch binding protein antibody
- G/T mismatch-binding protein antibody
see all
Images
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![Western blot - Anti-MSH6 antibody [44] (ab14204)](https://www.abcam.com/ps/products/14/ab14204/Images/ab14204-261775-ab14204KOWB.jpg)
Western blot - Anti-MSH6 antibody [44] (ab14204)Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: MSH6 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: A431 cell lysate (20 µg)
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)](https://www.abcam.com/ps/products/14/ab14204/Images/ab14204-284488-ICCGR262732101.jpg)
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).
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![Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [44] (ab14204)](https://www.abcam.com/ps/products/14/ab14204/Images/ab14204-284491-ICCGR262732101.jpg)
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).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [44] (ab14204)](https://www.abcam.com/ps/products/14/ab14204/Images/ab14204_1.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [44] (ab14204)ab14204 staining human colon carcinoma by IHC-P.
Protocols
References
This product has been referenced in:
- 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). Read more (PubMed: 29947891) »
