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Rabbit Recombinant Monoclonal MSH6 antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples.

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Images

Western blot - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (AB251498), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (AB251498), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (AB251498), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (AB251498), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (AB251498), expandable thumbnail

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBICC/IFFlow Cyt (Intra)IHC-P
Human
Tested
Tested
Tested
Tested
Tested

Tested
Tested

Species

Human

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

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      Target data

      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. Recruited on chromatin in G1 and early S phase via its PWWP domain that specifically binds trimethylated 'Lys-36' of histone H3 (H3K36me3): early recruitment to chromatin to be replicated allowing a quick identification of mismatch repair to initiate the DNA mismatch repair reaction.

      Alternative names

      DNA mismatch repair protein Msh6, hMSH6, G/T mismatch-binding protein, MutS protein homolog 6, MutS-alpha 160 kDa subunit, GTBP, GTMBP, p160, GTBP, MSH6

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      Rabbit Recombinant Monoclonal MSH6 antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples.

      Alternative names

      DNA mismatch repair protein Msh6, hMSH6, G/T mismatch-binding protein, MutS protein homolog 6, MutS-alpha 160 kDa subunit, GTBP, GTMBP, p160, GTBP, MSH6

      Key facts

      Isotype

      IgG

      Form

      Liquid

      Clonality

      Monoclonal

      Immunogen
      • The exact immunogen used to generate this antibody is proprietary information.
      Carrier free

      Yes

      Clone number

      EPR20316

      Purification technique

      Affinity purification Protein A

      Specificity

      This antibody recognizes isoforms 1 and 4 of MSH6.

      Concentration
      Loading...

      Storage

      Shipped at conditions

      Blue Ice

      Appropriate long-term storage conditions

      +4°C

      Storage information

      Do Not Freeze

      Notes

      ab251498 is the carrier-free version of Anti-MSH6 antibody [EPR20316] ab208940.

      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 monoclonal antibody, which offers several advantages including:

      • - High batch-to-batch consistency and reproducibility
      • - Improved sensitivity and specificity
      • - Long-term security of supply
      • - Animal-free batch production

      For more information, read more on recombinant antibodies.

      Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

      This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

      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.

      This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

      Supplementary info

      Activity summary

      MSH6 also known as MutS Homolog 6 is a DNA repair protein that plays a role in the mismatch repair (MMR) system. It has a molecular mass of approximately 136 kDa. MSH6 forms a heterodimer with MSH2 called MutSα and this complex identifies base-pair mismatches and insertion-deletion loops during DNA replication. It is expressed in various tissues throughout the body and high levels are often found in proliferative tissues where active DNA replication occurs.

      Biological function summary

      MSH6 functions as part of the MMR complex which is essential for maintaining genomic stability. The MutSα complex where MSH6 pairs with MSH2 operates along with other proteins in the MMR pathway to correct DNA replication errors. MSH6 is also known to interact with PCNA a DNA polymerase processivity factor which facilitates its role in the repair process.

      Pathways

      MSH6 participates prominently in the DNA mismatch repair pathway. This pathway is critical for correcting DNA errors and preventing mutations during replication. In association with MLH1-PMS2 (MutLα complex) MSH6 ensures that DNA integrity is preserved. Additionally MSH6 is involved in the base excision repair (BER) pathway where it collaborates with other repair proteins to fix small base lesions.

      Associated diseases and disorders

      MSH6 has a significant connection with Lynch syndrome also known as hereditary nonpolyposis colorectal cancer (HNPCC). This condition is characterized by germline mutations in MMR genes including MSH6 leading to increased cancer risk particularly in the colon. Moreover alterations in MSH6 can contribute to microsatellite instability a feature seen in certain types of endometrial cancer. Mutations in MSH2 often accompany MSH6 mutations in these disorders further impacting the MMR pathway's efficiency.

      Product promise

      We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

      In the unlikely event of one of our products not working as expected, you are covered by our product promise.

      Full details and terms and conditions can be found here:
      Terms & Conditions.

      9 product images

      • Western blot - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498), expandable thumbnail

        Western blot - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498)

        This data was developed using Anti-MSH6 antibody [EPR20316] ab208940, the same antibody clone in a different buffer formulation.

        Lane 1: Wild type HAP1 whole cell lysate (20 μg)

        Lane 2: MSH6 knockout HAP1 whole cell lysate (20 μg)

        Lane 3: HeLa whole cell lysate (20 μg)

        Lane 4: A431 whole cell lysate (20 μg)

        Lanes 1 - 4: Merged signal (red and green). Green - Anti-MSH6 antibody [EPR20316] ab208940 observed at 165 kDa. Red - loading control, ab18058, observed at 130 kDa.

        Anti-MSH6 antibody [EPR20316] ab208940 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 samples were used. Wild-type and MSH6 knockout samples were subjected to SDS-PAGE. Anti-MSH6 antibody [EPR20316] ab208940 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

        All lanes: Western blot - Anti-MSH6 antibody [EPR20316] (Anti-MSH6 antibody [EPR20316] ab208940)

        Predicted band size: 153 kDa

      • Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498), expandable thumbnail

        Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498)

        This data was developed using Anti-MSH6 antibody [EPR20316] ab208940, the same antibody clone in a different buffer formulation.

        Anti-MSH6 antibody [EPR20316] ab208940 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 Anti-MSH6 antibody [EPR20316] ab208940 at 1μg/ml and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed 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).

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498), expandable thumbnail

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498)

        This data was developed using Anti-MSH6 antibody [EPR20316] ab208940, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human endometrium tissue labeling MSH6 with Anti-MSH6 antibody [EPR20316] ab208940 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

        Nuclear staining on tumor cells of human endometrium is observed [PMID: 8942985].

        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.

        Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

      • Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498), expandable thumbnail

        Immunocytochemistry/ Immunofluorescence - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498)

        This data was developed using Anti-MSH6 antibody [EPR20316] ab208940, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling MSH6 with Anti-MSH6 antibody [EPR20316] ab208940 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green).

        Confocal image showing nuclear staining on HeLa cell line.

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

        Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498), expandable thumbnail

        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498)

        This data was developed using Anti-MSH6 antibody [EPR20316] ab208940, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling MSH6 with Anti-MSH6 antibody [EPR20316] ab208940 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

        Nuclear staining on tumor cells of human colon cancer is observed [PMID: 26315971].

        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.

        Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

      • Western blot - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498), expandable thumbnail

        Western blot - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498)

        This data was developed using Anti-MSH6 antibody [EPR20316] ab208940, the same antibody clone in a different buffer formulation.

        Blocking and dilution buffer: 5% NFDM/TBST.

        All lanes: Western blot - Anti-MSH6 antibody [EPR20316] (Anti-MSH6 antibody [EPR20316] ab208940) at 1/1000 dilution

        All lanes: Human testis tissue lysate at 20 µg

        Secondary

        All lanes: Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

        Predicted band size: 153 kDa

        Observed band size: 120 kDa

        Exposure time: 3min

      • Immunoprecipitation - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498), expandable thumbnail

        Immunoprecipitation - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498)

        This data was developed using Anti-MSH6 antibody [EPR20316] ab208940, the same antibody clone in a different buffer formulation.

        MSH6 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with Anti-MSH6 antibody [EPR20316] ab208940 at 1/40 dilution.

        Western blot was performed from the immunoprecipitate using Anti-MSH6 antibody [EPR20316] ab208940 at 1/1000 dilution.

        VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/1000 dilution.

        Lane 1: HeLa whole cell lysate, 10 μg (Input).

        Lane 2: Anti-MSH6 antibody [EPR20316] ab208940 IP in HeLa whole cell lysate.

        Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-MSH6 antibody [EPR20316] ab208940 in HeLa whole cell lysate.

        Blocking and dilution buffer: 5% NFDM/TBST.

        Exposure time: 8 seconds.

        The observed molecular weight corresponds to isoform 1 (160kDa) and isoform 4 (120kDa) which is consistent with the Uniprot annotation.

        All lanes: Immunoprecipitation - Anti-MSH6 antibody [EPR20316] (Anti-MSH6 antibody [EPR20316] ab208940)

        Predicted band size: 153 kDa

        Observed band size: 120 kDa, 160 kDa

      • Western blot - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498), expandable thumbnail

        Western blot - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498)

        This data was developed using Anti-MSH6 antibody [EPR20316] ab208940, the same antibody clone in a different buffer formulation.

        Blocking and dilution buffer: 5% NFDM/TBST.

        Exposure time: Lanes 1-3: 3 minutes; Lane 4: 15 seconds.

        This antibody recognizes isoforms 1 and 4 of MSH6. The observed molecular weight corresponds to isoform 1 (160kDa) and isoform 4 (120kDa) which is consistent with the Uniprot annotation.

        All lanes: Western blot - Anti-MSH6 antibody [EPR20316] (Anti-MSH6 antibody [EPR20316] ab208940) at 1/1000 dilution

        Lane 1: A431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

        Lane 2: SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

        Lane 3: HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

        Lane 4: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

        Secondary

        All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

        Predicted band size: 153 kDa

        Observed band size: 120 kDa, 160 kDa

      • Flow Cytometry (Intracellular) - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498), expandable thumbnail

        Flow Cytometry (Intracellular) - Anti-MSH6 antibody [EPR20316] - BSA and Azide free (ab251498)

        This data was developed using Anti-MSH6 antibody [EPR20316] ab208940, the same antibody clone in a different buffer formulation.

        Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling MSH6 with Anti-MSH6 antibody [EPR20316] ab208940 at 1/500 dilution (red) compared with a rabbit monoclonal IgG isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

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