153 A novel liquid nitrogen-free snap freezer Supplementary Figure 2. Influence of freezing rate on phosphoproteomic profiles Experimental design of the comparison of the effect of 5 different freezing rates, achieved using 3 different types of vials in 2 different coolants, on the phosphoproteome using cancer cell line HCT116. Aliquots of 300 µl HCT116 lysate, each corresponding to 300-400 µg of protein, were placed in three types of vials with different thermal conduction properties (polypropylene, aluminum and aluminum vials covered in paper tape) to influence their freezing rates. For each condition three vials were individually immersed in either LN2 or precooled isopentane for 1 minute and cooled to a temperature of -196 °C or -80 °C, respectively, using a stainless steel vial holder. The time to reach the goal temperature of -80 °C was registered. 5
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