PCODEPseudo Code
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#: 161-0375); bovine serum albumin (BSA) from Sigma-Aldrich, Co., MO, USA (PCode: 1001561418); 4',6-diamidino-2-phenylindole (DAPI); ethanol "99%" and [beta]-mercaptoethanol from Carl Roth GmbH + Co.
Total mixture volume was 200 uL in a well contained 140 uL mM HEPES (bioworld: cat#40820000-1) Tris (Invitrogen: cat# 15504-020) buffer of pH 7.4), 20 uL of enzyme (sigma Aldrich, C2624, PCode: 1001584424) (0.1 - 0.2 mg mL in deionized water), 20 uL (0.5 mg mL in DMSO) of test extract was mixed and incubated at 25o C for 15 minutes.
Rabbit colon total protein was extracted using radio immunoprecipitation assay (RIPA) lysis buffer (PCode: P0013C, Beyotime Institute of Biotechnology, Shanghai, China) consisting of 50 mM TRIS (pH 7.4), 150 mM NaCl, 1% Nonidet P (NP-40), 0.5% sodium deoxycholate, 0.1% SDS, and protease inhibitors such as sodium orthovanadate, sodium fluoride, and EDTA.