pro-q diamond phosphoprotein gel stain  (Thermo Fisher)

Journal: bioRxiv

Article Title: Spatially resolving how phosphorylation affects ß-cardiac myosin activity in porcine myofibril sarcomeres with single molecule resolution

doi: 10.1101/2024.06.27.600847

Figure Lengend Snippet: Protein was isolated from homogenized myofibrils and treated with PKA and MLCK and compared to untreated myofibrils. Myofibrils per condition were run on a 15% SDS PAGE gel (A) as well as a 4-20% gradient SDS-PAGE gel (B) to allow for better separation and resolution of the cMyBP-C band. Each gel was stained with Pro-Q Dimond Phosphoprotein Gel Stain (right) followed by Coomassie Blue stain (left) to quantify total protein. Densitometry analysis was performed on the cMyBP-C and RLC phosphorylation bands and normalized to the actin band of the respective Coomassie gel stain. (C) cMyBP-C phosphorylation per treatment group was calculated relative to the untreated group (N=1, n=2). (D) RLC phosphorylation per treatment group was calculated relative to the untreated group (N=1, n=3). Data shown as mean ± SEM. Student’s T-test was performed relative to untreated *p<0.05. Uncropped gels are provided in the supplementary material (Figure S2).

Article Snippet: Gels were stained with Pro-Q Diamond Phosphoprotein Gel Stain (ThermoFisher Scientific, P33300) to determine the phosphorylation levels of sarcomeric proteins.

Techniques: Isolation, SDS Page, Staining