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. Author manuscript; available in PMC: 2022 Jan 1.
Published in final edited form as: Methods Mol Biol. 2021;2259:25–45. doi: 10.1007/978-1-0716-1178-4_3

Fig. 1. Effect of fixations, protein extractions and dehydration of samples on protein yield.

Fig. 1.

GLU/PFA fixed samples (A) lysed with RIPA buffer (0.01% SDS) on ice or (B) lysed with RIPA buffer (2% SDS) and incubations at 100°C/20 min and 60°C/2h; (1) Unfixed lysates, (2) 4% PFA alone, (3) GLU (0.05%)/PFA, (4) GLU (0.01%)/PFA, (5) GLU (0.005%)/PFA; (C) PFA/DTBP fixed samples were lysed with RIPA buffer (2% SDS and 100 mM DTT) and incubations at 37°C/30 min, 100°C/20 min and 60°C/2h; (1) Unfixed lysates, (2) 4% PFA alone, (3) PFA/3mM DTBP, (4) PFA/5mM DTBP, (5) PFA/10mM DTBP. (D) PFA/5mM DTBP fixed samples extracted and loaded as in gel C; (1) Cells were fixed and lysed the same day; (2) Cells were fixed and kept in PBS for 24h before lysing; (3) Cells were fixed and kept dry for 24h before lysing; (4) Cells were fixed and kept in PBS for 72h before lysing; (5) Cells were fixed and kept dry for 72h before lysing. Stars indicate protein bands with decreased intensity. Gels are representative of 3 independent experiments (reproduced from ref. 26 with permission from Proteomics).