The High-Throughput Crystallization Screening Center performs crystal-growth screening experiments in 1536-well microassay plates for both soluble and membrane biological macromolecules. We set up the experiments with a predetermined set of crystallization cocktails, image them weekly over six weeks, and send you those images including SONICC and UV along with analysis software to view them. You send the sample, we do the rest.
- Sample requirements
500 μl of protein sample in a labeled 1.5 ml microcentrifuge tube. We make the following recommendations:
- Send only high purity, non-aggregated, stable protein samples.
- Include all chemicals required for protein stability.
- Use low (<25mM) buffer concentration. Avoid phosphate buffer (forms insoluble salts).
- Strive for 10 mg/ml protein concentration when solubility permits.
- Do not lyophilize your protein.
The current cost for 1,536 different chemical conditions in the soluble or membrane screen is $375 US. Users are billed after the sample has been set up and the experiment completed (~6 weeks). The service is for academic and other not-for-profit users only. If you are a commercial user, please visit our friends, ‘the structure people‘ at www.harkerbio.com.
- Sending your sample
Please send an email message to email@example.com. In this message, specify (i) the name of your institution, (ii) the number of protein samples you would like to assay, and (iii) for each protein specify whether you want the soluble (standard) or membrane protein crystallization screening assay.
Space will rapidly be reserved for you in the experimental queue, and a return message will provide you with (i) the date to send your sample(s), (ii) a Submission Form for tracking your protein, and (iii) an Order Form with payment options. Both forms must be completed and returned to firstname.lastname@example.org to finalize your reservation.
Once the paperwork is completed and your reservation is confirmed samples should be shipped to:
High-Throughput Crystallization Screening Center
Hauptman-Woodward Medical Research Institute
700 Ellicott Street
Buffalo, New York 14203, USA
Users must include a hard copy of the Sample Form along with the protein solution in a clearly labeled 1.5 ml tube placed inside of a protective container.
We make the following recommendations:
- Ship on Monday-Wednesday to reduce the risk of a weekend delivery delay.
- Use an insulated Styrofoam shipping container.
- Use dry ice for frozen samples.
- Use gel packs to maintain 4°C or 23°C sample temperatures.
- What you get
1,536 Microbatch-under-oil crystallization screening experiments are set up with your sample. For the soluble screen, the assay includes an incomplete factorial based PEG/Salt/Buffer screen (688 cocktails) and chemically complementary screens purchased from Hampton Research which includes: Crystal Screen HT, Index, PEGRx HT, SaltRx HT, PEG/Ion HT, Slice pH, Ionic liquid, Polymer Screen, Silver-bullets, and others (848 cocktails). The membrane screen, developed by the Malkowski lab, samples regions of detergent phase space particularly relevant to crystallization (1344 cocktails) with the Hampton Research PEGRx screen (96 cocktails), and the Molecular Dimensions MemGold screen (96 cocktails).
Email notification and immediate access to color images of your experimental outcomes as they are recorded: (i) cocktail only prior to adding sample (control), (ii) plus images taken 1 day, (iii) 1 week, (iv) 2 weeks, (v) 3 weeks, (vi) 4 weeks and (vii) 6 weeks after adding sample.
Email notification and access to images from a Formulatrix Rock Imager 1000 SONICC (to identify chiral crystals) and UV-TPEF (to determine if crystals are biological). These images are taken either at ~4 weeks (23°C samples) or ~6 weeks (4°C or 14°C samples). Actual read time may vary slightly and is dependent upon availability of the system. Thus, there are a total of nine sets of images recorded over a period of six weeks.
MacroScopeJ software (Java-based, multi-platform) to view, annotate and analyze images as well as associated chemical data.
Information will not be publicly disclosed regarding any sample received for crystallization trials without the expressed written consent of the investigator(s).
For information on the screens used and the Center the following articles may be of interest. If the high-throughput screening facility assists you in identifying initial crystallization conditions, please reference the appropriate one:
- Soluble Screen. Luft, J. R., Collins, R. J., Fehrman, N. A., Lauricella, A. M., Veatch, C. K. & DeTitta, G. T. (2003). A deliberate approach to screening for initial crystallization conditions of biological macromolecules. J. Struct. Biol. 142, 170-179.
- Membrane Screen. Koszelak-Rosenblum, M., Krol, A., Mozumdar, N., Wunsch, K., Ferin, A., Cook, E., Veatch, C. K., Nagel, R., Luft, J. R., DeTitta, G. T & Malkowski, M. G. (2009). Determination and application of empirically derived detergent phase boundaries to effectively crystallize membrane proteins. Protein Science 18, 1828-1839.
- Mailing list
We like to keep users up to date with new developments, e.g. improved cocktails, analysis software etc. If you would like to sign up for our mailing list to these notifications, please send a blank message to the following address: Announcementsemail@example.com
- Special requirements
We can often accommodate special requirements, please contact us at firstname.lastname@example.org.