Rabbit Recombinant Monoclonal MSH2 antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 1 publication.
IgG
Rabbit
pH: 7.2 - 7.4
Constituents: PBS
Liquid
Monoclonal
IP | WB | ICC/IF | Flow Cyt (Intra) | IHC-P | |
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Human | Tested | Expected | Tested | Tested | Tested |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
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Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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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.
DNA mismatch repair protein Msh2, hMSH2, MutS protein homolog 2, MSH2
Rabbit Recombinant Monoclonal MSH2 antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 1 publication.
DNA mismatch repair protein Msh2, hMSH2, MutS protein homolog 2, MSH2
IgG
Rabbit
pH: 7.2 - 7.4
Constituents: PBS
Liquid
Monoclonal
Yes
EPR21017-123
Affinity purification Protein A
Blue Ice
+4°C
Do Not Freeze
ab228334 is the carrier-free version of Anti-MSH2 antibody [EPR21017-123] ab227941.
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:
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.
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.
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.
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.
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.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling MSH2 with Anti-MSH2 antibody [EPR21017-123] 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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MSH2 antibody [EPR21017-123] ab227941).
Immunofluorescent analysis of 100% methanol-fixed A-375 (human malignant melanoma cell line) cells labeling MSH2 with Anti-MSH2 antibody [EPR21017-123] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MSH2 antibody [EPR21017-123] ab227941).
MSH2 was immunoprecipitated from 0.35 mg of A-375 (human malignant melanoma cell line) lysate with Anti-MSH2 antibody [EPR21017-123] ab227941 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-MSH2 antibody [EPR21017-123] 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: Anti-MSH2 antibody [EPR21017-123] ab227941 IP in A-375 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-MSH2 antibody [EPR21017-123] ab227941 in A-375 whole cell lysate.
Exposure time: 1 second.
Blocking and dilution buffer concentration: 5% NFDM/TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MSH2 antibody [EPR21017-123] ab227941).
All lanes: Immunoprecipitation - Anti-MSH2 antibody [EPR21017-123] (Anti-MSH2 antibody [EPR21017-123] ab227941)
Developed using the ECL technique.
Predicted band size: 105 kDa
Observed band size: 105 kDa
Immunofluorescent analysis of 100% methanol-fixed A549 (human lung carcinoma cell line) cells labeling MSH2 with Anti-MSH2 antibody [EPR21017-123] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MSH2 antibody [EPR21017-123] ab227941).
Immunohistochemical analysis of paraffin-embedded human testis tissue labeling MSH2 with Anti-MSH2 antibody [EPR21017-123] 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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MSH2 antibody [EPR21017-123] ab227941).
Overlay histogram showing HAP1 wildtype (green line) and HAP1-MSH2 knockout cells (red line) stained with Anti-MSH2 antibody [EPR21017-123] 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 Anti-MSH2 antibody [EPR21017-123] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MSH2 antibody [EPR21017-123] ab227941).
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized A-375 (human malignant melanoma cell line) cell line labeling MSH2 with Anti-MSH2 antibody [EPR21017-123] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-MSH2 antibody [EPR21017-123] ab227941).
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