Tetragonal Lysozyme crystallization
published: 14 Nov 2006 (14:24)
Chicken egg white Lysozyme CrystallizationThere are many different recipes for chicken egg white Lysozyme crystallization. The main common feature, of these crystallization techniques, described below is their simplicity.
Please note: in order to protect your samples from bacterial or fungal infection you must add about 0.02% (w/v) Sodium azide in all solutions you use. Filter all solutions with a micro filter of 0.22 microns.
There are two major crystallization techniques, used not only for Lysozyme crystallization, but for almost all proteins are: hanging drop and sitting drop. The main difference in these two techniques is the volume of the protein drop in which crystals are grow. For hanging drop the average volume is about 1-5 microliters, whereas for sitting drop 10-20 microliters is usually used.
Lysozyme crystallization procedure 1The crystals obtained from these crystallization conditions are more suitable for non-frozen X-Ray data collection.
Note: Last trials shows that these crystallization conditions can be cryoprotective, especially when the drop with Lysozyme crystals partially dry out.
Lysozyme crystallization procedure 2The crystals obtained from these crystallization conditions are more suitable for low temperature X-Ray data collection, because of presence of cryo-protective compounds in the Lysozyme crystallization conditions.
Lysozyme crystalsUsually, chicken egg white Lysozyme crystals are grown in two different crystal forms. Furthermore, the appearance of these crystal forms is not related with Lysozyme crystallization conditions, but in our experiments we obtain only tetragonal Lysozyme crystals.
Tetragonal Lysozyme crystals
For more information about all crystallographic techniques you can read in the Guide to the Protein crystallography, from Protein crystallization to Structure refinement. Furthermore, you can look through one of the biggest crystallographic bulletin board to see how all scientist deal with these tasks.
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