MSH2 antibody | knockout validation | Abcam ab70270
DOI
//dx.doi.org/10.13070/ko.en.6.1625
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).

MSH2 antibody | knockout validation | Abcam ab70270 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: Abcam

Antibody: MSH2

Catalog number ab70270

Summary: Rabbit polyclonal MSH2 antibody validated for WB, IP, IHC, ICC/IF and tested in Human and Mouse.

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-rabbit IgG, NA934V, 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