Western Blot Analysis of Protein Extracts

Protein extracts were prepared as described above and incubated for 5 min. at 100 °C. Different amounts of proteins were loaded on 4–20% gradient acrylamide gel and transferred onto PVDF membrane. After incubation with corresponding antibodies, the protein analysis was performed using ECL-PLUS kit (GE-Healtcare) as described before and VersaDoc apparatus (Bio-Rad) as previously described^{35 (link)}. Primary antibodies used: Anti-FLAG -HRP conjugated antibody (abcam; 1:10000), Anti Actin (abcam 1:500) and Anti RAD-51 (1:1000).

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Germoglio M., Valenti A., Gallo I., Forenza C., Santonicola P., Silva N, & Adamo A. (2020). In vivo analysis of FANCD2 recruitment at meiotic DNA breaks in Caenorhabditis elegans. Scientific Reports, 10, 103.

Publication 2020

VersaDoc apparatus Anti-FLAG -HRP conjugated antibody Anti Actin

Acrylamide Actin Antibodies Antibody Membrane Protein Pvdf

Corresponding Organization :

Other organizations : Institute of Biosciences and Bioresources, National Research Council, Masaryk University

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Variable analysis

independent variables

Different amounts of proteins loaded on 4–20% gradient acrylamide gel

dependent variables

Protein analysis performed using ECL-PLUS kit (GE-Healtcare) and VersaDoc apparatus (Bio-Rad)

control variables

Protein extracts prepared as described above
Protein extracts incubated for 5 min. at 100 °C
Proteins transferred onto PVDF membrane
Primary antibodies used: Anti-FLAG -HRP conjugated antibody (abcam; 1:10000), Anti Actin (abcam 1:500) and Anti RAD-51 (1:1000)

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