1) Buffer preparation: Prepare 10L HBSS balanced salt solution with sterile water.
HBSS balanced salt solution formula: 400mg/L KCl, 60mg/L KH2PO4, 350mg/L NaHCO3, 8000mg/L NaCl, 121mg/L Na2HPO4·12H2O, 1000mg/L Glucose.
2) Weigh 1gm of Recombinant Trypsin (cell culture grade), dissolve and clarify with HBSS balanced salt solution to obtain Recombinant Trypsin solution with an enzyme activity concentration of 500 USP units/ml-1500 USP units/ml or mass concentration of 0.1mg/ml-0.3mg/ml. (The concentration of 500 USP units/ml fits most cell detachment while primary cell, tissue etc. may need the concentration of 1500 USP units/ml.)
3) Adjust the recombinant trypsin solution’s PH to 7.2~8.0 with NaOH solution and hydrochloric acid, than sterile filter with a 0.22 μm filter membrane to obtain the recombinant trypsin digestion solution.
1) This Recombinant Trypsin digestion solution contains EDTA, so attention should be paid to its suitability for different cells.
2) This Recombinant Trypsin digestion solution contains no bacteriostatic agents. Special attention should be paid to the aseptic operation during use to avoid the contamination of the digestion solution by microorganisms.
3) It is suggested to divide this Recombinant Trypsin digestion solution into 100ml/vial which can be stored stably at -15~-20℃ for 12 months. It is not suitable for long-term storage at 4℃, and repeated freezing and thawing should be avoided.
