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Perparing Iodide or Cesium derivatives
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for a SIR, SAD, or SIRAS experiment
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5-amino-2,4,6-triiodoisophthalic acid (I3C)
Adapted from Beck et al., 2008, A magic triangle for experimental phasing of macromolecules. Acta Cryst vol D64, 1179-1182. and Beck et al., 2009, How to get the magic triangle and the MAD triangle into your protein crystal. Acta Cryst vol F65, 1068-70.
I3C has the advantage of 3 iodide atoms per molecule for additional phasing power.
See Tobias Beck's web page for additional information.
(1) I3C is located in room 219A, in the drawer below the balance. The drawer is labeled "Heavy atom ends". In a small brown glass bottle with red lid.
It was ordered from Sigma http://www.sigmaaldrich.com/catalog/search/ProductDetail/ALDRICH/444367
(2) Weigh 0.03g I3C (30 mg)
(3) Dissolve I3C in 59 uL of 2.0 M lithium hydroxide. (Makes 1.0M I3C solution) (I3C=556 g/mol) as recommended by Tobias.
(4) Mix 3.5 microliters I3C solution with 6.5 microliters reservoir and 3.5 microliters 100% glycerol stock for cryoprotection. This makes approximately 260 mM I3C, final concentration. Tobias recommends up to 500mM concentration.
(more or less glycerol may be required for cryoprotection)
(5) Soak crystal 5 minutes in the cryo-I3C solution.
(6) Mount crystal on cold stream
As a positive control, I3C was used to derivatize proteinase K. You can download the XDS formatted structure factors or the Scalepack formatted structure factors and log file.
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