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Protein labeling

AT Andrea Testa
MD Mirco Dindo
AR Aleksander A. Rebane
BN Babak Nasouri
RS Robert W. Style
RG Ramin Golestanian
ED Eric R. Dufresne
PL Paola Laurino
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l-lactate dehydrogenase and Jack bean urease were tagged using Alexa Fluor 594 and 488 Microscale Protein Labeling Kit respectively, using the manufacturer’s protocol but repeating the dialysis step three times to remove the free dye in the protein solution. The BSA in Fig. 1a was tagged with Alexa Fluor 647 Microscale Protein Labeling Kit using the same purification procedure. The reaction between SNARF-1 and BSA was carried out in potassium phosphate 0.1 M pH 7.0 in presence of sodium bicarbonate 0.1 M for at least 1 h at room temperature. Tagged proteins were further purified using membrane dialysis to reduce the free probe in solution. To determine the protein concentration of LDH and urease we used ε280nm = 43,680 M−1 cm−1 and ε280nm = 54,165 M−1cm−1, respectively (https://web.expasy.org/protparam/).

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