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Before sending sample for crystallization trial

 Prior to start a new crystallization project, you first need to purify the protein in relatively large quantities (few milligrams). High purity and homogeneity of the sample are crucial for the crystallization to be successful. Dynamic light scattering (DLS) may be used to characterize the polydispersity of the sample (the presence of different oligomeric forms or even aggregates of the protein in solution) after concentrating the sample, and thermofluor for characterizing the stability of the protein in different buffers and in the presence of different ligands. We recommend also to characterize the protein by MS (MW and protein identity). Several other biophysical methods like CD spectroscopy may also be used if further characterization of the protein is required. If you need help or advice about sample preparation and quality control, please contact the SPC facility .

 
 
 
Required sample volume

Sitting and hanging drops

Please use this formula to calculate the minimum volume required to carry on your crystallization experiments

[ (total protein volume needed per condition) x 96 condition x (number of plate requested)] + 15% + 10ul)

  • Example 1 (1 drop per condition) =>  (100nl protein per well x 96 conditions x 1 plate + 15%) + 10ul = 21.04ul total => round up => 22ul minimum.
  • Example 2 (2 drops per condition, different drop ratio) => ([200nl protein + 100nl protein]each well x 96 conditions x 5 plates + 15%) + 10ul = 175.6ul total => round up => 176ul minimum.

      Find details about how to prepare a seed stock solution
here.
 

Sample delivery

Sample requirements:

  • Provide in 0.5ml microcentrifuge tube(s)
  • Clearly label everything using a permanent marker.
  • Please avoid excess use of scotch tape or parafilm.
  • The samples should be delivered to the SPC.

Customs clearance?
Users outside EU should provide all information necessary about their sample(s), with the shipment, directly to the customs (charges may apply). The SPC will not set up crystallization trial of sample(s) which have been delayed more than 2 days due to custom clearance.

 


Delivery address

Sample Prepration and Characterization facility
EMBL (c/o DESY)
Notkestrasse 85, Build. 25a
22607 Hamburg
Germany