Team:UCLA/Notebook/Recombinant Expression/2 June 2015
- Experimental Design Brainstorm
- Tamura-GFP Hydrogel assay
- Negative control - sfGFP without Tammy.
- Experimental protein - Tamura
- Assay for binding potential to doped silk solution.
- Used stripping buffers to assay for agitation. (detergents, alkalinity, ionic strength, pH).
- Differing variables: Concentration of Dope, concentration of Tamura in hydrogel.
- Temperature based gradient on hydrogel stability.
- Assay for flourescence using microplane reading, distribution using fluorescence intensity (read into measuring this quality).
- Tamura-GFP Hydrogel assay
Characterization of Tamura -
- SDS-PAGe gels, Western Blot to assay degradation.
- CD, IR Spectrum, Fluorescence Intensity of purified protein and compare that with protein concentration - relative to that metric (ug).
- Cry - EM for silk Tamura fibers - make sure that we don't mess with the fibers.
- Gel meshes and pore size using EM.
- Tamura-ABD Assays
- Incubate in albumin containing solution for set time, measure the rate of release of albumin within the hydrogel. Hydrogel with ABD-Tam, and ABD out.
- CD, IR, before and post vortex of the solution.
- Tamura-ABD Assays
- Flourescence following washes of Tamura.