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Rabbit Recombinant Monoclonal MSH2 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 12 publications.

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH2 antibody [EPR21017-123] (AB227941), expandable thumbnail
  • Western blot - Anti-MSH2 antibody [EPR21017-123] (AB227941), expandable thumbnail
  • Immunoprecipitation - Anti-MSH2 antibody [EPR21017-123] (AB227941), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-MSH2 antibody [EPR21017-123] (AB227941), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-MSH2 antibody [EPR21017-123] (AB227941), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

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

1/30

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

1/1000

Notes

-

Tested
Tested

Species

Human

Dilution info

1/100

Notes

-

Tested
Tested

Species

Human

Dilution info

1 µg/mL

Notes

MeOH fixationis recommended

Tested
Tested

Species

Human

Dilution info

1/8000

Notes

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

Associated Products

Select an associated product type

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Target data

Function

Component of the post-replicative DNA mismatch repair system (MMR). Forms two different heterodimers: MutS alpha (MSH2-MSH6 heterodimer) and MutS beta (MSH2-MSH3 heterodimer) which binds to DNA mismatches thereby initiating DNA repair. When bound, heterodimers bend the DNA helix and shields approximately 20 base pairs. MutS alpha recognizes single base mismatches and dinucleotide insertion-deletion loops (IDL) in the DNA. MutS beta recognizes larger insertion-deletion loops up to 13 nucleotides long. After mismatch binding, MutS alpha or beta 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. Recruits DNA helicase MCM9 to chromatin which unwinds the mismatch containing DNA strand (PubMed:26300262). 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. In melanocytes may modulate both UV-B-induced cell cycle regulation and apoptosis.

Alternative names

DNA mismatch repair protein Msh2, hMSH2, MutS protein homolog 2, MSH2

Recommended products

Rabbit Recombinant Monoclonal MSH2 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 12 publications.

Alternative names

DNA mismatch repair protein Msh2, hMSH2, MutS protein homolog 2, MSH2

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

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

EPR21017-123

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

Notes

To see more of the key markers and tools you need to study the hallmarks of cancer, including genome instability and mutation, please visit the following page.

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

MSH2 also known as MutS Homolog 2 is a human protein with a molecular weight of approximately 100 kDa. It is an important component of the DNA mismatch repair system and plays an essential role in maintaining genomic stability by recognizing and repairing mismatched nucleotides during DNA replication. Expression of the MSH2 protein occurs broadly in dividing cells across various tissues with notable presence in tissues with high proliferation rates such as the colon and the endometrium. Additionally detection and quantification of MSH2 are often performed using methodologies such as MSH2 ELISA which aids in the assessment of its expression levels in different biological samples.

Biological function summary

Components are identified in mismatch repair where MSH2 forms a heterodimer with MSH6 known as the MutSα complex or with MSH3 known as the MutSβ complex. This heterodimerization is critical for the initial steps in the recognition and binding of mismatch errors on the DNA strand. MSH2 complex formation enables it to scan the DNA for errors facilitating the recruitment of additional repair proteins. The activity of MSH2 in these complexes is important in preserving the fidelity of genetic information and prevents mutations that could lead to genomic instability.

Pathways

MSH2 operates within the DNA damage response and repair pathways. The protein is a core component of the mismatch repair pathway which corrects DNA replication errors that elude proofreading activity of DNA polymerases. It interacts with other proteins such as MLH1 and PMS2 forming a synergistic function that amplifies the capacity to recognize and initiate repair of mismatches. The pathway involving MSH2 not only repairs mismatched bases but also plays a role in cell cycle control checkpoints and apoptosis evidencing its pivotal role in maintaining cell cycle integrity.

Associated diseases and disorders

Studies show that MSH2 is strongly associated with Lynch syndrome an autosomal dominant inherited condition that increases the risk of colorectal cancer. Mutations in the MSH2 gene impair its mismatch repair function and lead to microsatellite instability a hallmark of cancer cells in this disorder. Furthermore alterations in the MSH2 protein also relate to glioblastomas with correlations observed between MSH2 expression levels and tumor progression. These conditions exemplify the important role of MSH2 and its interaction with other DNA repair proteins in preventing cancerous developments.

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:
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9 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH2 antibody [EPR21017-123] (ab227941), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH2 antibody [EPR21017-123] (ab227941)

    Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling MSH2 with ab227941 at 1/8000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in para-carcinoma colonic epithelium (image B) or stromal cells (both image A and B) and loss of expression in the paired colon cancer (image A) (PMID: 24710284). Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

  • Western blot - Anti-MSH2 antibody [EPR21017-123] (ab227941), expandable thumbnail

    Western blot - Anti-MSH2 antibody [EPR21017-123] (ab227941)

    Exposure times: Lane 1: 100 seconds; Lane 2: 3 minutes; Lane 3: 10 seconds; Lane 4: 1 minute; Lane 5: 15 seconds.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

    All lanes: Western blot - Anti-MSH2 antibody [EPR21017-123] (ab227941) at 1/1000 dilution

    Lane 1: HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

    Lane 2: A549 (human lung carcinoma epithelial cell), whole cell lysate at 10 µg

    Lane 3: Human fetal heart lysate at 10 µg

    Lane 4: Human tonsil lysate at 10 µg

    Lane 5: A-375 (human malignant melanoma epithelial cell), whole cell lysate at 10 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

    Developed using the ECL technique.

    Predicted band size: 105 kDa

    Observed band size: 104 kDa

  • Immunoprecipitation - Anti-MSH2 antibody [EPR21017-123] (ab227941), expandable thumbnail

    Immunoprecipitation - Anti-MSH2 antibody [EPR21017-123] (ab227941)

    MSH2 was immunoprecipitated from 0.35 mg of A-375 (human malignant melanoma cell line) lysate with ab227941 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab227941 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution.

    Lane 1: A-375 whole cell lysate 10 μg (Input).

    Lane 2: ab227941 IP in A-375 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab227941 in A-375 whole cell lysate.

    Exposure time: 1 second.

    Blocking and dilution buffer concentration: 5% NFDM/TBST.

    All lanes: Immunoprecipitation - Anti-MSH2 antibody [EPR21017-123] (ab227941)

    Developed using the ECL technique.

    Predicted band size: 105 kDa

    Observed band size: 105 kDa

  • Flow Cytometry (Intracellular) - Anti-MSH2 antibody [EPR21017-123] (ab227941), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-MSH2 antibody [EPR21017-123] (ab227941)

    Overlay histogram showing HAP1 wildtype (green line) and HAP1-MSH2 knockout cells (red line) stained with ab227941. The cells were fixed with 80% methanol (5 min) (left panel) or 4% formaldehyde (10 min) (right panel) , and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by ab227941 for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) presorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution for 30 min at 22°C.

    A rabbit IgG isotype control antibody (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-MSH2 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

    Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

    Note: We recommend fixing cells using MeOHinstead of PFA toget optimal results.

  • Immunocytochemistry/ Immunofluorescence - Anti-MSH2 antibody [EPR21017-123] (ab227941), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-MSH2 antibody [EPR21017-123] (ab227941)

    Immunofluorescent analysis of 100% methanol-fixed A549 (human lung carcinoma cell line) cells labeling MSH2 with ab227941 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Nuclear staining in A549 cell line is shown.

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

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH2 antibody [EPR21017-123] (ab227941), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH2 antibody [EPR21017-123] (ab227941)

    Immunohistochemical analysis of paraffin-embedded human testis tissue labeling MSH2 with ab227941 at 1/8000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in human testis was observed (PMID: 10029069). Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunocytochemistry/ Immunofluorescence - Anti-MSH2 antibody [EPR21017-123] (ab227941), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-MSH2 antibody [EPR21017-123] (ab227941)

    Immunofluorescent analysis of 100% methanol-fixed A-375 (human malignant melanoma cell line) cells labeling MSH2 with ab227941 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Nuclear staining in A-375 cell line is shown.

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

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

  • Flow Cytometry (Intracellular) - Anti-MSH2 antibody [EPR21017-123] (ab227941), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-MSH2 antibody [EPR21017-123] (ab227941)

    Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized A-375 (humanmalignant melanoma cell line) cell line labeling MSH2 with ab227941 at 1/600 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - 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 H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Western blot - Anti-MSH2 antibody [EPR21017-123] (ab227941), expandable thumbnail

    Western blot - Anti-MSH2 antibody [EPR21017-123] (ab227941)

    Western blot: Anti-MSH2 antibody [EPR21017-123] (ab227941) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab227941 was shown to bind specifically to MSH2. A band was observed at 104 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in MSH2 knockout cell line. To generate this image, wild-type and MSH2 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    All lanes: Western blot - Anti-MSH2 antibody [EPR21017-123] (ab227941) at 1/1000 dilution

    Lane 1: Wild-type HCT 116 cell lysate at 20 µg

    Lane 2: MSH2 knockout HCT 116 cell lysate at 20 µg

    Lane 3: A-375 cell lysate at 20 µg

    Observed band size: 104 kDa

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