MSH3 antibody | knockout validation | Santa Cruz sc-271079
DOI
//dx.doi.org/10.13070/ko.en.6.1626
Date
2016-09-22

This is a knockout-validated antibody summary, based on the publication "A MutSβ-Dependent Contribution of MutSα to Repeat Expansions in Fragile X Premutation Mice?", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

MSH3 antibody | knockout validation | Santa Cruz sc-271079 figure 1
Figure 1. Equivalent amounts of total protein from brain, testes and ovary and nuclear extracts from liver of three 6-month old WT, Msh3-/- and Msh6-/- and Msh2-/- mice were subjected to electrophoresis and western blotting using the MSH2, MSH3 and MSH6 antibodies, from figure 4 [1].
Antibody information

Company: Santa Cruz

Antibody: sc-271079

Catalog number: sc-271079

Summary: MSH3 Antibody (C-8) is a high quality mouse monoclonal recommended for detecting MSH3 of human origin by WB, IP, IF, IHC(P) and ELISA.

Validation Method

Western blot

Sample

Flash frozen brain, liver, testes and ovary of 6-month old mice, homogenized using a tissue homogenizer (Precellys 24, Bertin Technologies, Berlin, Germany) with T-PER protein extraction reagent (Pierce Biotechnology, Inc, Rockford, IL) supplemented with complete, Mini, EDTA-free protease inhibitor cocktail (Roche Applied Science, Indianapolis, IN).

Nuclear extracts of liver proteins: NE-PER Nuclear and cytoplasmic extraction reagents (Pierce Biotechnology, Inc, Rockford, IL).

The protein concentrations were determined using a Bio-Rad protein assay kit (Bio-Rad, Hercules, CA). Proteins were heated for 10 minutes at 70°C in LDS-Sample Buffer (Life Technologies, Grand Island, NY), resolved by electrophoresis on either 3–8% NuPAGE Novex Tris-Acetate gels (Life Technologies) or 4–12% NuPAGE Novex Tris-Bis gels (Life Technologies) and transferred to nitrocellulose membranes using the iBlot transfer apparatus (Life Technologies).

Blocking agent

Membranes were blocked for one hour at room temperature in 5% ECL Prime blocking agent (GE Healthcare Bio-Sciences) in TBST

Primary incubation

Incubated overnight at 4°C at 1:10000 dilution.

Secondary incubation

anti-mouse IgG, NA931V GE Healthcare Bio-Sciences, at a dilution of 1:5000.

Detection

ECL Prime detection reagent (GE Healthcare Bio-Sciences), Fluorchem M imaging system (Proteinsimple, Santa Clara, CA).

References
  1. Zhao X, Lokanga R, Allette K, Gazy I, Wu D, Usdin K. A MutSβ-Dependent Contribution of MutSα to Repeat Expansions in Fragile X Premutation Mice?. PLoS Genet. 2016;12:e1006190 pubmed publisher