Overview

  • Product name

    Anti-MSH6 antibody [SP93] - BSA and Azide free
    See all MSH6 primary antibodies
  • Description

    Rabbit monoclonal [SP93] to MSH6 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human MSH6 aa 350-450 (internal sequence). The exact sequence is proprietary.
    Database link: P52701

  • Epitope

    Internal region
  • Positive control

    • WB: Wild-type HAP1 cell lysate FC: HeLa cells IHC-P: Human rectal carcinoma, colon, and colon carcinoma tissues; Mouse colon tissue
  • General notes

    Ab238805 is the carrier-free version of ab99889. 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.

    ab238805 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A/G purified
  • Purification notes

    Purified from TCS by protein A/G.
  • Clonality

    Monoclonal
  • Clone number

    SP93
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab238805 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 Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.

Antigen retrieval: Boil tissue section in 1 mM EDTA, pH 8.0 for 10 minutes followed by cooling at room temperature for 20 minutes. Primary antibody incubation: 30 minutes at room temperature.

ICC/IF Use at an assay dependent concentration.
Flow Cyt 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

  • 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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse colon
    tissue sections labeling MSH6 with ab99889 at 1/100 dilution (1.0 µg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Sporadically nuclear staining on mouse colon, performed on a Leica Biosystems BOND™ RX instrument.
    The section was incubated with ab99889 for 30 mins at room temperature. This image was generated using ab99889, the same clone, but with a different buffer formulation.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon carcinoma tissue sections labeling MSH6 with ab99889 at 1/100 dilution (1.0 µg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Nuclear staining on human colon carcinoma, performed on a Leica Biosystems BOND™ RX instrument.
    The section was incubated with ab99889 for 30 mins at room temperature. This image was generated using ab99889, the same clone, but with a different buffer formulation.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue sections labeling MSH6 with ab99889 at 1/100 dilution (1.0 μg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Sporadically nuclear staining on human colon, performed on a Leica Biosystems BOND™ RX instrument.
    The section was incubated with ab99889 for 30 mins at room temperature. This image was generated using ab99889, the same clone, but with a different buffer formulation.

  • 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 - ab99889 observed at 160 kDa. Red - loading control, ab18058, observed at 124 kDa.

    ab99889 was shown to specifically react with MSH6 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed in MSH6 knockout samples. Wild-type and MSH6 knockout samples were subjected to SDS-PAGE. ab99889 and ab18058 (loading control to Vinculin) were diluted at 1 μg/ml and 1/10 000 respectively and incubated overnight at 4°C. 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 1/10 000 dilution for 1 h at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA and sodium azide (ab99889).

  • ab99889 staining MSH6 in wild-type HAP1 cells (top panel) and MSH6 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), 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 ab99889 at 1/250 dilution and ab195889 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 Rabbit IgG (Alexa Fluor® 488) (ab150081) 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab99889).

  • Flow cytometry analysis of HeLa (human cervix adenocarcinoma) labeling MSH6 with purified ab99889 at 1/20 dilution (5.05 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as a secondary antibody. Isotypecontrol - Rabbit monoclonal IgG (ab172730) (black). Unlableled control - Unlabelled cells (blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab99889).

  • Staining of MSH6 in a formalin fixed, paraffin embedded Human rectal carcinoma tissue using ab99889 at a dilution of 1/100.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab99889).

  • ab99889 staining MSH6 in HeLa cells. The cells were fixed with 100% methanol (5min), 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 ab99889 at 1/250 dilution and ab195889 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 Rabbit IgG (Alexa Fluor® 488) (ab150081) 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab99889).

References

ab238805 has not yet been referenced specifically in any publications.

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