Anti-MSH2 antibody [3A2B8C] (ab52266)
Key features and details
- Mouse monoclonal [3A2B8C] to MSH2
- Suitable for: Flow Cyt, ICC/IF, WB, IHC-P, IP
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-MSH2 antibody [3A2B8C]
See all MSH2 primary antibodies -
Description
Mouse monoclonal [3A2B8C] to MSH2 -
Host species
Mouse -
Tested applications
Suitable for: Flow Cyt, ICC/IF, WB, IHC-P, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Ni-NTA purified recombinant human MSH2 expressed in E. Coli strain BL21 (DE3).
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Positive control
- WB: HeLa, A549, A431 and HEK-293 cell lysates. ICC/IF: HeLa cells. Flow cyt: HeLa cells. IHC-P: Human rectum carcinoma tissue. IP: HEK-293 whole cell lysate.
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General 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.
This product was changed from ascites to supernatant. Lot no’s high than GR128648-25 are from Tissue Culture Supernatant
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
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Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
Preservative: 0.05% Sodium azide
Constituent: PBS -
Concentration information loading...
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Purity
Protein G purified -
Purification notes
Purified from tissue culture supernatant. -
Clonality
Monoclonal -
Clone number
3A2B8C -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab52266 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 |
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Flow Cyt |
Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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ICC/IF | (1) |
1/200 - 1/1000.
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WB | (2) |
1/500 - 1/2000. Detects a band of approximately 98 kDa (predicted molecular weight: 105 kDa).
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IHC-P |
1/200 - 1/1000.
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IP |
Use a concentration of 5 µg/ml.
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Notes |
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Flow Cyt
Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/200 - 1/1000. |
WB
1/500 - 1/2000. Detects a band of approximately 98 kDa (predicted molecular weight: 105 kDa). |
IHC-P
1/200 - 1/1000. |
IP
Use a concentration of 5 µg/ml. |
Target
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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. 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. -
Tissue specificity
Ubiquitously expressed. -
Involvement in disease
Defects in MSH2 are the cause of hereditary non-polyposis colorectal cancer type 1 (HNPCC1) [MIM:120435]. 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. The term "suspected HNPCC" or "incomplete HNPCC" can be used to describe families who do not or only partially fulfill the Amsterdam criteria, but in whom a genetic basis for colon cancer is strongly suspected. MSH2 mutations may predispose to hematological malignancies and multiple cafe-au-lait spots.
Defects in MSH2 are a cause of Muir-Torre syndrome (MuToS) [MIM:158320]; also abbreviated MTS. MuToS is a rare autosomal dominant disorder characterized by sebaceous neoplasms and visceral malignancy.
Defects in MSH2 are a cause of susceptibility to endometrial cancer (ENDMC) [MIM:608089].
Defects in MSH2 are a cause of hereditary non-polyposis colorectal cancer type 8 (HNPCC8) [MIM:613244]. HNPCC is a disease associated with marked increase in cancer susceptibility. It is characterized by a familial predisposition to early-onset colorectal carcinoma (CRC) and extra-colonic tumors 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. Clinically, HNPCC is often divided into two subgroups. Type I is characterized by hereditary predisposition to colorectal cancer, a young age of onset, and carcinoma observed in the proximal colon. Type II is characterized by 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. The term 'suspected HNPCC' or 'incomplete HNPCC' can be used to describe families who do not or only partially fulfill the Amsterdam criteria, but in whom a genetic basis for colon cancer is strongly suspected. Note=HNPCC8 results from heterozygous deletion of 3-prime exons of EPCAM and intergenic regions directly upstream of MSH2, resulting in transcriptional read-through and epigenetic silencing of MSH2 in tissues expressing EPCAM. -
Sequence similarities
Belongs to the DNA mismatch repair mutS family. -
Post-translational
modificationsPhosphorylated by PRKCZ, which may prevent MutS alpha degradation by the ubiquitin-proteasome pathway.
Phosphorylated upon DNA damage, probably by ATM or ATR. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 4436 Human
- Omim: 609309 Human
- SwissProt: P43246 Human
- Unigene: 597656 Human
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Alternative names
- BAT26 antibody
- COCA 1 antibody
- COCA1 antibody
see all
Images
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All lanes : Anti-MSH2 antibody [3A2B8C] (ab52266) at 1/2000 dilution
Lane 1 : Cell lysates prepared from human Hela cells
Lane 2 : Cell lysates prepared from A549 cells
Lane 3 : Cell lysates prepared from human A431 cells
Lane 4 : Cell lysates prepared from HEK293 cells
Lysates/proteins at 100 µg per lane.
Secondary
All lanes : HRP-conjugated Goat polyclonal to mouse IgG
Predicted band size: 105 kDa -
ab52266 (1/200) detecting MSH2 in HeLa cells (green). Cells were fixed in methanol (-20'C, 10min) and counterstained with DAPI in order to highlight the nucleus. Please refer to abreview for further experimental details.
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ab52266 at 1/1000 dilution staining MSH2 in human Hela cells by Immunocytochemistry/ Immunofluorescence. An Alexa Fluor® 488 conjugated Goat polyclonal to mouse IgG1 was used as secondary antibody. The primary antibody shows green staining in image whilst actin filaments were stained red with Alexa Fluor® 555 phalloidin.
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Overlay histogram showing HeLa cells stained with ab52266 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52266, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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MSH2 was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Mouse monoclonal to MSH2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab52266.
Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/10,000 dilution.
Band: 105kDa; MSH2 -
Immunohistochemical analysis of paraffin-embedded human rectum carcinoma tissue, showing nuclear and cytoplasmic localisation, using ab52266 at a dilution of 1/200 - 1/1000 with DAB staining.
Protocols
Datasheets and documents
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Datasheet download
References (30)
ab52266 has been referenced in 30 publications.
- Guan B et al. Identification of Germline Mutations in Upper Tract Urothelial Carcinoma With Suspected Lynch Syndrome. Front Oncol 12:774202 (2022). PubMed: 35372080
- Gurumurthy RK et al. Patient-derived and mouse endo-ectocervical organoid generation, genetic manipulation and applications to model infection. Nat Protoc 17:1658-1690 (2022). PubMed: 35546639
- Hsueh CY et al. Fusobacterium nucleatum impairs DNA mismatch repair and stability in patients with squamous cell carcinoma of the head and neck. Cancer 128:3170-3184 (2022). PubMed: 35789992
- Franck S et al. MSH2 knock-down shows CTG repeat stability and concomitant upstream demethylation at the DMPK locus in myotonic dystrophy type 1 human embryonic stem cells. Hum Mol Genet 29:3566-3577 (2021). PubMed: 33242073
- Ji X et al. PD-L1, PARP1, and MMRs as potential therapeutic biomarkers for neuroendocrine cervical cancer. Cancer Med 10:4743-4751 (2021). PubMed: 34076351