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In partnership with
Oxford Cryosystems
for North America, Canada
and South America

 

Discover - Five great ideas for optimizing your protein crystals.
MRC MAXI Optimization Plate - ideal for optimization of crystal hits.
MEET - us in person at BCA

Five great ideas for optimizing your protein crystals.

Optimization of a protein crystallization hit can be challenging to say the least. Nothing is as frustrating as getting an initial hit only to find the crystal does not diffract as well as hoped, or is twinned etc.

Optimization of a crystallization condition requires a refinement of the existing conditions or the addition of extra components. Here are five great tips for making optimization a little easier:

  1. Do: check the exact formulation of the ‘hit’ including how it was buffered and how the buffer was titrated. There are many different ways to make up a crystallization ‘hit, and the best way is to contact the manufacturer to check you are using the correct titrants etc. We always give our customers the information required to make up their ‘hits’ including lot numbers and suppliers of chemicals as well as titrants, to give you the best possible start with your optimization.

  2. Test: different temperatures. Try setting-up
    your crystallization at different temperatures. Temperature has been shown to be a very significant variable when it comes to obtaining good quality protein crystals. If your initial
    set-up was at room temperature, try 4
    degrees and vice versa. Or if you have
    incubators- for example compact Bench-top incubators can be set at different temperatures
    so you can easily test a range of temperatures.

  3. Change: drop volume. Sometimes bigger drops can give bigger crystals. But you need to be aware about the scale-up issue on moving from nanolitre to microliter and 96-well plates to 24-well plates. Scaling is not a linear process. As scale and volume increase, the ratio between the surface area and
    volume decreases.The MRC MAXI 48-well optimization plate
    was specifically developed by crystallographers at the MRC in Cambridge to help alleviate this scale-up issue. You can easily scale-up your drop size either by robot or by hand and yet keep the environment close to what you had in a 96-well plate. So you get larger volume drops for optimization, but use less of your screen and space in the incubator or lab.

    Get a free MRC 48-well MAXI Plate by e-mailing us.

  4. Try: seeding. Seeding is used to improve the size of crystals, provide more consistent results, speed-up results if nucleation is slow and help obtain a wider range of crystal forms by seeding into totally different precipitants. MicroSeed Beads, for example, are used to generate microseeds – (microscopic crystals that have been ‘crushed-up’ into fragments); It is easier to add onto an already existing nucleus than create one de novo. Here are some seeding exercises to try in your lab.

  5. Add: Nucleants. If you cannot bear to crush your precious crystals, then adding a nucleant is the way to go. Nucleants, and these can include hair and skin, promote nucleation. Naomi’s nucleants, for example, work not only by promoting crystal growth in the optimization screen, but can also promote a ‘more-ordered’ crystal growth leading to potentially improved diffraction.

MEET- us in person at:
BCA Spring Meeting

At The University of Loughborough
7 - 10 April, 2014.

Come and meet Dr. Ali Abdul-Gader who will be happy to answer any of your protein crystallography questions.


Did you miss the last eNEWS with exciting news about..

Crystallizing complexes - Oligomers are increasingly important targets!
Editorial - Exploring more crystallization space.
Want to find more hits? - see the advantages of fluorescent labeling...


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