Tris hcl buffer solution

Deionized water and Tris-HCl buffer solution (pH 8.0, 1 M) were purchased from Sangon Biotech (Shanghai, China). Lithium chloride was purchased from Shanghai Aladdin Bio-Chem Technology (Shanghai, China). T4 DNA ligase and 10 × T4 DNA ligase reaction buffer (400 mM Tris-HCl, 100 mM MgCl2, 100 mM DTT, 5 mM ATP, pH 7.8) were purchased from Sangon Biotech (Shanghai, China).

Spermine tetrahydrochloride, silver nitrate (AgNO₃), sodium borohydride (NaBH₄), adenosine 5′-triphosphate disodium salt hydrate (ATP), protein kinase A (PKA, from bovine heart), and dithiothreitol (DTT) were purchased from Sigma–Aldrich (Shanghai, China). Dihydrochloride hydrate (H-89) was obtained from J&K Scientific Ltd. (Shanghai, China). Protein kinase B (PKB) was purchased from Bio-Techne China Co., Ltd. (Shanghai, China). Forskolin, 3-isobutyl-1-methylxanthine (IBMX), and Tris-HCl buffer solution (pH 7.5) were purchased from Sangon Biotech (Shanghai, China). Substrate peptide for PKA (TAMRA-LRRASLG, 98%), and phosphorylated substrate peptide were purchased from GL Biochem (Shanghai, China). Magnesium chloride (MgCl₂), sodium chloride (NaCl), ethylenediaminetetraacetic acid (EDTA), glycerol, and dimethyl sulfoxide (DMSO) were obtained from Shanghai Chemicals Ltd. (Shanghai, China). The human cervical carcinoma cells (HeLa) were purchased from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). All other reagents were of A.R. grade and used as received without further purification. Ultrapure water with a resistivity of 18.2 MΩ·cm was used.