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Low myocardial protein kinase G activity in heart failure with preserved ejection fraction
Protein quantity was determined by Bradford assay (Bio-Rad Laboratories, 5000006) and equal amounts of protein were denatured at 95 C in Laemmli buffer, resolved on SDS-PAGE and transferred to a polyvinylidene fluoride membrane (Sigma, GE10600023)
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This 1:1 ratio prevents peptide precipitation (which occurs in overly concentrated solutions) while avoiding excessive dilution that wastes refrigerator space and increases contamination risk across multiple draws
It affects the red blood cells, causing episodes of 'sickling', which produce episodes of pain and other symptoms
