Proteins were extracted using RIPA (Radioimmunoprecipitation Assay) buffer containing 1% proteinase inhibitor (Thermo Scientific) and then quantified using Bradford reagent (Protein Assay Dye Reagent Concentrate, BIO-RAD) with bovine serum albumin as a standard.
After two washes in ethanol-based buffer (Buffer RPE, Qiagen), RNA were eluted in RNase-free water and then quantified by optical density measurement using Nanodrop ND-100 spectrophotometer (Nanodrop Technologies, Wilmington, DE).
All synthesized sequences were purified by HPLC and then quantified.
Resulting purified protein was dissolved in assay buffer (62.5 mM Na HEPES, 62.5 mM NaCl, 62.5 mM NaF, 6.25 mM Na pyrophosphate, 1.25 mM EDTA, 1.25 mM EGTA, 1 mM dithiothreitol, 1 mM PMSF, 5 µg/mL SBTI and 1 µM pepstatin A) and then quantified using the bicinchoninic assay.
The enzymatic reaction was purified using the MinElute Reaction Cleanup Kit and the DNA recovered with two 20 ul buffer EB elutions, then quantified as described above.
It then produces images for melanin, blood, and collagen, which are then quantified and analytically evaluated.
For normalization of the radioactivity with total amount of protein, the cells pellets obtained as above were lysed with lysis buffer and then the protein was quantified.
The membrane was washed 5 times with tris buffer, and then specific bands were quantified using a ChemiImager System (Alpha Innotech Corporation).
Soluble and insoluble fractions were then quantified by Bradford reagent.
