Visualizing Recombinant and Patient Derived Poly(GA) Proteins
Corresponding Organization : Mayo Clinic in Florida
Other organizations : Mayo Clinic, WinnMed, Boston University
Protocol cited in 1 other protocol
Variable analysis
- Dilution of recombinant GST or GST-(GA)50 proteins to 1 µg/µl in 20 mM Tris–HCl, pH 7.5, 50 mM KCl, 10 mM MgCl2
- Incubation of the samples at 30 °C for 6 h
- Dilution of the samples to 0.1 µg/µl by reaction buffer
- Use of mouse monoclonal anti-GST antibody (1:20) as primary antibody for immuno-EM analysis
- Use of goat anti-mouse IgG conjugated with 6 nm colloidal gold particles (1:20) as secondary antibody for immuno-EM analysis
- Use of rabbit polyclonal anti-poly(GA) antibody (1:50) as primary antibody for immuno-EM analysis of patient samples
- Use of goat anti-rabbit IgG conjugated with 18 nm colloidal gold particles (1:20) as secondary antibody for immuno-EM analysis of patient samples
- Filamentous structure of recombinant GST-(GA)50 proteins
- Filamentous structure of poly(GA) proteins in c9FTD/ALS patient samples
- Recombinant GST protein as a control for the GST-(GA)50 protein
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