Instructions:

- In each eppendorf, there is 3.7mg of HS dleu -roll. - The folded -roll domain consists of two -sheet faces, each with eight solvent exposed residues. These leucine-rich surfaces impart a hydrophobic driving force for self-assembly. We genetically fused an -helical leucine zipper domain (H) capable of forming tetrameric coiled-coil bundles and a randomly coiled hydrophilic linker (S) to the peptide and the resulting chimeric protein self-assembles into a hydrogel only in calcium rich environments.- Without calcium, the helices form tetrameric coiled-coil bundles, but the -roll domains remain unstructured, delocalizing the leucine-rich patches and prohibiting supramolecular self-assembly. After calcium binding, the -roll folds and the leucine-rich faces are exposed creating a hydrophobic driving force for oligomerization, resulting in the hydrogel network formation.- 618ul of diluted protein samples were lyophilized in 5mM Tris. In order to achieve a 6wt% hydrogel formation in 50mM Tris Buffer (pH 7.4), the powder in each tube should be re-suspended in 61.8ul of DI water (1/10 of the original volume).