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After 72 h, the resulting solution was freeze-dried by using low-temperature freeze dryers, yielding P1(1.23 g) as a yellow powder

2D crystallization The crystallization procedure was performed as described 15 with a different crystallization buffer: 25 mM Tris-HCl at pH 7.4, 20% glycerol, 0.1 mM EDTA, 100 mM KCl, 1 mM GSH, and 50 mM CaCl 2 and crystallization was performed at 30 C

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However, the compound became widely available for routine use in the treatment of B12 deficiency in the early 1950s (8)

The mean background value for each channel was further used in automated image analysis