MSH2 Recombinant Rabbit Monoclonal Antibody [JE28-48]
Catalog# ET7110-50
MSH2 Recombinant Rabbit Monoclonal Antibody [JE28-48]
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WB
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IHC-P
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FC
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Human
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unconjugated
Overview
Product Name
MSH2 Recombinant Rabbit Monoclonal Antibody [JE28-48]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human MSH2 aa 1-100 (N terminal).
Species Reactivity
Human
Validated Applications
WB, IHC-P, FC
Molecular Weight
Predicted band size: 105 kDa
Positive Control
HL-60 cell lysates, Wild-type SCC7 whole cell lysates, human colon carcinoma tissue, Hela.
Conjugation
unconjugated
Clone Number
JE28-48
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:500-1:2,000
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IHC-P
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1:50-1:200
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FC
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1:50-1:100
Target
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. 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.
Background References
1. Takeda T. et. al. Synchronous endometrial and ovarian cancer in Lynch syndrome with a MSH2 germline mutation: A case report. Mol Clin Oncol. 2018 Nov;9(5):479-484.
2. Arshita N. et. al. The Tendency of Having MSH2 and MSH6 Microsatellite Instability among Clinicopathological Features in Patients with Colorectal Cancer. Asian Pac J Cancer Prev. 2018 Nov 29;19(11):3147-3152.
Sequence Similarity
Belongs to the DNA mismatch repair MutS family.
Tissue Specificity
Ubiquitously expressed.
Post-translational Modification
Phosphorylated by PRKCZ, which may prevent MutS alpha degradation by the ubiquitin-proteasome pathway.
Subcellular Location
Nucleus, chromosome.
UNIPROT
Synonyms
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
ExpandImages
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☑ Knockout (KO)
All lanes: Western blot analysis of MSH2 with anti-MSH2 antibody [JE28-48] (ET7110-50) at 1/1,000 dilution.
Lane 1: Wild-type SCC7 whole cell lysate (20 µg).
Lane 2: MSH2 knockout SCC7 whole cell lysate (20 µg).
ET7110-50 was shown to specifically react with MSH2 in wild-type SCC7 cells. No band was observed when MSH2 knockout sample was tested. Wild-type and MSH2 knockout samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET7110-50, 1/1,000) and Loading control antibody(Rabbit anti-HSP90, ET1605-56, 1/10,000)was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of MSH2 on HL-60 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET7110-50, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
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Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-MSH2 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7110-50, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Flow cytometric analysis of MSH2 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7110-50, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"