Lysozyme crystallization recipe for stuffed

printed: 14 November 2006 (13:24)

Chicken egg white-colored Lysozyme Crystallization

There are various recipes for chicken egg white-colored Lysozyme crystallization. The primary common feature, of those crystallization techniques, described here is their simplicity.

Please be aware: to be able to safeguard your samples from microbial or yeast infection you have to add about .02% (w/v) Sodium azide in most solutions you utilize. Filter all solutions having a micro filter of .22 microns.

There's two major crystallization techniques, used not just for Lysozyme crystallization. but for most proteins are: hanging drop and sitting drop. The primary improvement in both of these techniques is the level of the protein stop by which crystals are grow. For hanging drop the typical volume is all about 1-5 microliters, whereas for sitting drop 10-20 microliters is generally used.

Lysozyme crystallization procedure 1

Urates acquired from all of these crystallization the weather is more appropriate for non-frozen X-Ray data collection.

1. Prepare 75mg/ml sample of Lysozyme in .1 M NaAc pH 4.8.
2. Well solution includes 1ml of 6.5% (w/v) NaCl and .1 M NaAc pH 4.8.
3. Cover slip drop contains: 5-7 microliters of Lysozyme solution and 5-3 microliters of well solution for as many as 10 microliters.
4. Seal a protective cover slip with grease or vacuum lubricant over each well.
5. Crystals should grow within 1-a couple of days.

It may be beneficial to put together some crystallization trays with variation of protein/well solutions with variation of Lysozyme concentration between 50 mg/ml and 75 mg/ml.

Note: Last trials implies that these crystallization conditions could be cryoprotective, particularly when the drop with Lysozyme crystals partly dry up.

Lysozyme crystallization procedure 2

Urates acquired from all of these crystallization the weather is more appropriate for cold X-Ray data collection, due to existence of cryo-protective compounds within the Lysozyme crystallization conditions.

1. Prepare 75mg/ml sample of Lysozyme in .1 M NaAc pH 4.8.
2. Well solution includes 1ml of 10% (w/v) NaCl, .1M NaAc pH 4.8 and 25% (v/v) Ethylene Glycol.
3. Cover slip drop of contains: 6-7 microliters of Lysozyme solution and 4-3 microliters of well solution for any total drop size 10 microliters.
4. Seal a protective cover slip with grease or vacuum lubricant over each well.
5. Crystals should grow within 1-a couple of days and be prepared for freezing.

If how big crystals isn't appropriate for more analysis, it may be beneficial to put together some crystallization trays with variation of protein/well solutions with variation of Lysozyme concentration between 50 mg/ml and 75 mg/ml.

Lysozyme crystals

Usually, chicken egg white-colored Lysozyme crystals are grown in 2 different very forms. In addition, the look of these very forms isn't related to Lysozyme crystallization conditions, however in our experiments we have only tetragonal Lysozyme crystals.

Tetragonal Lysozyme crystals

Photography of Tetragonal Lysozyme crystals.