To begin, prepare an ECF-5 self-assembling peptide solution at a concentration of 10 milligrams per milliliter using a suitable buffer. Sonicate the mixture to dissolve the peptide, and store it at four degrees Celsius. Prepare a 50 milligrams per milliliter calcium chloride solution using deionized water.
Add 40 microliters of calcium chloride solution to 460 microliters of the ECF-5 peptide solution. Vortex to mix thoroughly, and incubate the mixture at room temperature for over two hours. Rapidly introduce one milliliter of PBS into the dimethyl sulfoxide or DMSO solubilized peptide.
Vortex thoroughly and incubate at room temperature. After five minutes, add 500 microliters of PBS, and let the mixture stand for 15 minutes. Then discard the supernatant and retain the bottom gel.
The ECF-5 peptide displayed glutathione-like reducing properties and self-assembly capability, forming a solid gel within minutes when the concentration exceeded 1%The ECF-5 hydrogel exhibited semi-solid properties when prepared via pH response, metal edition, or solvent exchange.
