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add 10ul of thrombin resin Mix resin with protein by pipetting Put the tubes thermomixer set @ 25C, 600rpm Time course: 0, 2, 4, 20hr At each time point, take the corresponding tube, spin down the reaction for 2-3 m @ 5000rpm Transfer 10ul of the supernatant from each tube to be analyzed on gel Mix the resin with the rest of the reaction again by pipetting After collecting samples for all time points, transfer the rest of the digestion mix to a fresh tube (should have ~60ul or 6ug left) Purification of cleaved protein by Glutathione resin GenScript Glutathione Sepharose 4B (cat # L00206, storage buffer 1x PBS with 20% ethanol) 1

Nazir S, Dhull SB, Sharma V (2025) Extraction and characterization of omega fatty acids (as fixed oil) from various plants seeds (flaxseeds, chia seeds, chilli seeds, basil seeds, quince seeds)