Crystallization for Academic, Government, and Not-for-Profit Institutes
Academic, government, and non-profit research institute users are enabled by NIH R24 GM124135 (all samples) and NSF 2029943 (for SARS-CoV-2 samples), as well as generous historical support from NIH. Industry users should make use of our alternate screening pathway. Samples are screened against 1,536 different crystallization conditions. 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, 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). Typically the experimental error for delivery of cocktails and/or protein is < 5% (most often the error is < 1%) and we use control experiments to rapidly identify and address any issues that arise.
Subsidized costs: Our academic, government and non-profit services are subsidized by NIH and NSF. Fee-for-service is $400 per experimental screen (billed after sample setup). Users are provided with all images, metadata and software to review their crystallization screening images.
Sample requirement: Currently, we require 500 μl of protein sample in a labeled 1.5 ml microcentrifuge tube. We recommend that you send only high purity, non-aggregated, stable samples including all chemicals required for stability. To ensure that the crystallization conditions dominate use low (<25mM) buffer concentration. Avoid phosphate buffer (which forms insoluble salts). A recommended concentration is 10 mg/ml where solubility permits but both lower and significantly higher concentrations have proved successful. We do not recommend lyophilizing your protein.
The steps to make use of the Crystallization Center
1. Make a reservation
Crystallization Screening is run with monthly deadlines and dates. To reserve a spot in the crystallization screening queue, please check those dates and email firstname.lastname@example.org by the reservation deadline.
Include the following information in your reservation email:
- User name and contact information
- PI/Institution name and contact information
- Number of samples you will be sending
- Approximate date you will be shipping the sample(s)
You will receive an email confirmation from us for your reservation.
2. Fill out and submit an online submission form
Click on “Submission Form” to fill this out with contact details, sample information, and payment. A pdf copy will be generated. A copy of this should be sent with the sample.
3. Ship your sample
Please include the pdf form generated when the submission form is filled out. Samples should be shipped in a clearly labeled 1.5 mL tube places inside of a protective container. Ensure the tube is sealed and protected against any potential leaks. An insulated Styrofoam shipping container is recommended with dry ice for frozen samples or gel packs to maintain 4°C or 23°C sample temperature.
We recommend shipping overnight Monday – Wednesday to reduce risk of a weekend delivery delay.
The HWI National Crystallization Center is active and able to support your scientific efforts, both on COVID-19 related projects and the crystallization of other targets. We’ve scheduled the soluble screen for September, and anticipate both a soluble and membrane screening run for October.
September 2021 Soluble
- Reservation Deadline: Fri September 17th
- Package Acceptance: Tues September 21st to Thurs September 30th
- Plate Setup:
- Tues September 21st – Thurs September 23rd
- Mon September 27th – Thurs September 30th
Note: Samples are not set up on Fridays.
Hauptman-Woodward Medical Research Institute
700 Ellicott Street
Buffalo, NY 14203-1102, USA
p: +1 716-898-8649
What you get:
1,536 microbatch-under-oil crystallization screening experiments set up with your sample.
Email notification and access to brightfield images of your experimental outcomes as they are recorded:
cocktail only prior to adding sample (control)
weeks 1, 2, 3, 4 & 6
Email notification and access to images from a Formulatrix Rock Imager 1000 SHG (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) – you choose the protocol.
Actual read time may vary slightly and is dependent upon the availability of the system.
A total of nine sets of images recorded over the 6 week experimental duration.
MacroScopeJ software (Java-based, multi-platform) to view, annotate, and analyze images as well as associated chemical data.
Typically the experimental error for delivery of cocktails and/or protein is < 5% (most often the error is < 1%) and we use control experiments to rapidly identify and address any issues that arise.
Our philosophy: We provide capabilities at the highest standards for quality, reliability, efficiency, and productivity while providing confidentiality and expert support for challenging experiments. We want you to be successful and help that by providing cost-effective access to world-class research capabilities and maintaining competitiveness.
Confidentiality: Information will not be publicly disclosed regarding any sample received for crystallization trials without the express written consent of the investigator(s). Crystallization images obtained at the Crystallization Center for academic, government, and non-profit laboratories may be used to support the scientific research of the Center, as well as in publications. Images will be de-identified from sample and user information unless the consent of the investigator is obtained.
Acknowledgments: Your use of the Crystallization Center is supported by federal funding. If the Center assists you in identifying initial crystallization conditions, we ask that you reference the appropriate publication below. They enable us to track our performance, demonstrate our role in the community to our funders, and continuously improve our processes and therefore your success.
- 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. We are also open to collaborative studies to enhance crystallization success.
High-Throughput Crystallization Screening Center
Hauptman-Woodward Medical Research Institute
700 Ellicott Street
Buffalo, NY 14203