Protein Detection via Western Blotting

Cell lysis and Western blotting were performed as described previously [31 (link)] The following antibodies were used: a polyclonal rabbit anti-Hsp104 antibody (Thermo Scientific Project NJ1240S), anti-Pgk1 monoclonal antibody (Molecular Probes, Carlsbad, CA, USA), and rabbit anti-Hsp70 antibody (Enzo SPA 757). HPR conjugated goat anti-rabbit or goat anti-mouse secondary antibodies (Thermo Fisher, Waltham, MA, USA) and SuperSignal Plus Chemiluminescent Substrate (Thermo Scientific) were used to develop the Western blot. Amersham Imager 600 was used to scan the blot and the intensities were quantified using the ImageJ-win64.

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Stanford K.E., Zhao X., Kim N., Masison D.C, & Greene L.E. (2023). Overexpression of Hsp104 by Causing Dissolution of the Prion Seeds Cures the Yeast [PSI+] Prion. International Journal of Molecular Sciences, 24(13), 10833.

Publication 2023

rabbit anti-Hsp70 antibody SuperSignal Plus Chemiluminescent Substrate Amersham Imager

Anti antibodies Anti antibody Antibodies Cell Goat Hsp70 Molecular probes Mouse Rabbit Scan Western blot

Corresponding Organization : National Institutes of Health

Other organizations : National Institute of Diabetes and Digestive and Kidney Diseases

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Hsp104 protein levels
Pgk1 protein levels
Hsp70 protein levels

control variables

Cell lysis and Western blotting procedures as described previously [31]

controls

Positive control: None mentioned
Negative control: None mentioned

Annotations

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