製品の概要

  • 製品名Anti-MSH2 antibody [3A2B8C]
    MSH2 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [3A2B8C] to MSH2
  • アプリケーション適用あり: Flow Cyt, ICC/IF, IHC-Fr, WB, IHC-P, ELISA, IPmore details
  • 種交差性
    交差種: Human, Monkey
  • 免疫原

    Ni-NTA purified recombinant human MSH2 expressed in E. Coli strain BL21 (DE3).

  • ポジティブ・コントロール
    • A431 cell lysate; human rectum carcinoma tissue.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.
IHC-Fr 1/200 - 1/1000.
WB 1/500 - 1/2000. Detects a band of approximately 98 kDa (predicted molecular weight: 105 kDa).
IHC-P Use at an assay dependent concentration.
ELISA 1/10000.
IP Use a concentration of 5 µg/ml.

ターゲット情報

  • 機能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.
  • 組織特異性Ubiquitously expressed.
  • 関連疾患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.
  • 配列類似性Belongs to the DNA mismatch repair mutS family.
  • 翻訳後修飾Phosphorylated by PRKCZ, which may prevent MutS alpha degradation by the ubiquitin-proteasome pathway.
    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • 細胞内局在Nucleus.
  • Information by UniProt
  • 参照データベース
  • 別名
    • BAT26 antibody
    • COCA 1 antibody
    • COCA1 antibody
    • DNA mismatch repair protein Msh2 antibody
    • FCC 1 antibody
    • FCC1 antibody
    • hMSH2 antibody
    • HNPCC 1 antibody
    • HNPCC antibody
    • HNPCC1 antibody
    • LCFS2 antibody
    • MSH 2 antibody
    • Msh2 antibody
    • MSH2_HUMAN antibody
    • MutS homolog 2 antibody
    • MutS homolog 2 colon cancer nonpolyposis type 1 antibody
    • MutS protein homolog 2 antibody
    see all

Anti-MSH2 antibody [3A2B8C] 画像

  • 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.

    See Abreview

  • 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
    HRP-conjugated Goat polyclonal to mouse IgG

    Predicted band size : 105 kDa
  • 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.

  • 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.

  • 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.

Anti-MSH2 antibody [3A2B8C] (ab52266) 使用論文

This product has been referenced in:
  • Nguyen HT  et al. Human embryonic stem cells show low-grade microsatellite instability. Mol Hum Reprod 20:981-9 (2014). Read more (PubMed: 25082980) »
  • Sirbu BM  et al. Identification of proteins at active, stalled, and collapsed replication forks using isolation of proteins on nascent DNA (iPOND) coupled with mass spectrometry. J Biol Chem 288:31458-67 (2013). WB . Read more (PubMed: 24047897) »

See all 5 Publications for this product

Product Wall

Application Western blot
Sample Human Cell lysate - whole cell (HeLa)
Gel Running Conditions Reduced Denaturing (4-12%)
Loading amount 30 µg
Specification HeLa
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 37°C
Username

Abcam user community

Verified customer

投稿 Jul 13 2015

Thank you for your enquiry regarding ab52266 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody. Additionally, thank you for supplying an image, as this has...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (iPS cells)
Loading amount 20 µg
Specification iPS cells
Gel Running Conditions Reduced Denaturing (4-12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Elisabetta Soragni

Verified customer

投稿 Apr 08 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Methanol
Permeabilization Yes - 0.5% Triton-X100 in PBS
Username

Dr. Kirk McManus

Verified customer

投稿 Nov 30 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"