Hello, I want to extract myosin from chicken breast. The operation is as follows: (1) the miced meat was washed with buffer A (0.1 M NaCl, 10 mM phosphate buffer，2 mM MgCl2, 1 mM EDTA, pH 7) for 3 times to obtain myofibrillar protein as a sediment; (2) the sediment was homogenized with Guba-Straub solution (0.3 M KCl, 0.1 M KH2PO4, 50 mM K2HPO4, 1 mM EDTA, 4 mM sodium pyrophosphate, pH 6.5) for 20 min at 4 ℃. However, the solution became so viscous that I couldn't gain the supernatant through centrifugation, even though the speed was up to 13,000 rpm for 12 min (5000 g for 12 min in the literature Food Chemistry 2018，242, 22–28). Could anyone tell me that how to handle this situation? Thank you very much for your nice suggestions.
Yan Li
Pay attention to the concentration of EDTA and the formation of its chelate complex with protein. It increases the viscosity of the solution as it binds protein molecules. It may be possible to reduce its concentration and remove inorganic components.