To begin the pH response method for hydrogel formation, add five milligrams of ECF-5 peptides to 400 microliters of deionized water, and sonicate it at 40 kilohertz for 30 minutes. Add 40 microliters of filtered one-molar sodium hydroxide to the peptide solution, and vortex to mix thoroughly. Continue sonication for 15 more minutes until the solution is completely clarified.
Next, add 60 microliters of hydrochloric acid to the prepared solution. After vortexing the mixture, let it stand at room temperature for over 30 minutes to facilitate hydrogel formation.