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Today's objective is to tag our BSA proteins with our dye. The dye we are using is 5(6)-Carboxynaphthofluorescein, N-succinimidyl ester and has been obtained from Marker Gene Technologies Inc. This dye is able to tag unprotonated lysine residues; for this reason the first step of our protocol is to increase the pH of our solutions (BSA/HCl control, 70 ratio, 166 ratio) so that it reaches a pH of 8.5 (because the lysine pKa = 8.95 for the amino group); to do so, we are going to add some buffer (prepared on 8/2/12) to the solution until the pH reaches 8.5 (we are checking the pH using a SB70P pHmeter).
Then, because the dye isn't water soluble, we have to prepare a satured DMSO solution; we are going to dissolve the dye (whose mass has been determined on 8/2/12) into as few DMSO as possible. To finish, we'll leave the reaction occur overnight; the next step will be to perform a dialysis to remove the excess of dye.
When we added the dye solution into our solutions, we observed an immediate color change.