Haynes Lab:Notebook/Engineering PC-TFs/2014/10/13

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Summary RFP Bradford Plate reader measurements RFP signal Followed the fluorescent protein assay: http://openwetware.org/wiki/Haynes:ExtFluorProtein Used 100 μL per well Results: U2OS BL09 = 181 Mock Transfection = 25 Bradford assay Followed Bradford Assay protocol: http://openwetware.org/wiki/Haynes:Bradford Used 5 μL for each BL09 sample OD 590 readings: Standard 1 (0 μg) = 0.276 Standard 2 (1 μg) = 0.234 Standard 3 (2 μg) = 0.394 Standard 4 (4 μg) = 0.443 Standard 5 (8 μg) = 0.644 Standard 6 (16 μg) = 0.829 U2OS BL09 = 0.313 Mock Transfection = 0.302 Standard curve Final results: Sample Protein μg/μL Protein*100 μL Raw Fluor. (100 uL) Fluor/ μg protein U2OS BL09 0.765 76.50 181 2.365929 Mock Transfection 0.464 46.45 25 0.538235