| Service items |
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CF-01 SDS-PAGE |
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Use precast gradient gel (4-12%) for sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) to verify purity of the submitted samples. |
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CF-02 UV scan & Ａ260/Ａ280 |
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UV-Vis was used to monitor protein quality. The ratio of absorbance at 260 nm versus 280 nm is commonly used to assess DNA contamination of protein solutions. A ratio of more than 0.8 is indicative of DNA contamination. |
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CF-03 Tycho NT.6 |
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Tycho NT.6 is a label-free Differential Scanning Fluorimeter (DSF). Upon heating samples from 35 to 95 ºC, it measures the ratio of the fluorescence intensities at 350 nm and 330 nm, which detects the environmental changes of the tryptophan and tyrosine residues. This technique is applicable for studies of thermal denaturation of proteins. |
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CF-04 Far-UV CD spectroscopy |
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Far-UV circular dichroism (CD) spectroscopy analyzes protein secondary structure contents and thermal stabilities. A six-well automated sample changer is available for simultaneous data collection. |
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CF-05 DSF Tm screen |
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Differential scanning fluorimetry (DSF) uses the Real-time Polymerase Chain Reaction (PCR) platform to screen for optimal sample conditions based on protein thermal stability readouts. By using the Solubility & Stability Screen 2™ kit in a 96-well format, a variety of combinations of pH and salt concentrations can be tested simultaneously. As a rule of sum, a higher Tm value increases the success rate in crystallization, better NMR spectral quality and cryo-EM sample integrity. |
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CF-06 AUC (SV mode) |
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Analytical ultracentrifuge (AUC) monitors particle sedimentation in solution. It is applicable to proteins, nucleic acids, carbohydrates and polymers. Sedimentation velocity (SV) mode is used to determine molecular weights (hence, oligomeric states), particle size and shape (conformations). |
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CF-07 SEC |
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High-resolution analytical size exclusion columns (GE 10/300) with three different matrices (Superdex 75 increase, Superdex 200 increase, Superose 6 increase) can be chosen for size exclusion chromatography (SEC) analysis to determine protein purity/homogeneity and oligomeric states. A baseline of the observed elution peaks is the prerequisite for reliable SEC-MALS analysis. |
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CF-08 SEC-MALS |
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Size exclusion chromatography-coupled multi-angle light scattering (SEC-MALS) is used to determine absolute molar mass of large molecules. The system is coupled with four on-line detectors: laser light scattering (miniDAWN TREOS, Wyatt), quasi-elastic light scattering (QELS, Wyatt), refraction index (Optilab T-rEX, Wyatt), and UV absorption detection. MALS combines the precise measurements of light scattering intensity and concentrations for a rigorous calculation of solution-property molecular weights—making no assumptions of conformation or shape, and no perturbations to the sample. In addition, QELS module can further provide simultaneous hydrodynamic radius (Rh) measurements in the same scattering volume viewed by the MALS detectors. SEC-MALS can be used to determine the oligomeric state of a target protein, quantify protein aggregation, and determine the stoichiometry of a protein conjugate. |
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CF-09 AF4-MALS |
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Asymmetric flow field-flow fractionation (AF4) can be used to analyze samples such as proteins, antibodies, micro-liposomes, exsosomes, extracellular vesicles, polymers, and nano particles. Traditional colloidal filtration chromatography techniques cannot effectively separate very large particles, such as aggregating precipitants, samples that adsorb to colloids, molecular weight of the molecule to be measured outside the column separation range, etc. AF4 can separate particle sizes up to several microns, and their corresponding molecular mass values can be determined downstream by MALS. The detected molecular mass range can reach the ninth power of ten Dalton. |
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PX-01 Robotic crystallization screening |
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Robotic screening of crystallization conditions is conducted by using an automated liquid dispenser (Phoenix/RE) and various crystallization plates.
Details of screening suites available: Please see the attachment 1.
Details of application for crystallization screening: Please see the attachment 2.
If you have any questions, please contact Dr. Kai-Fa Huang (02-27855696#4010; huangkf@gate.sinica.edu.tw).
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PX-02 Incubators for crystallization plates |
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We provide incubators with various temperatures for crystal growth. |
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PX-03 Crystal inspection with UV-imaging system |
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We provide a real-time UV-imaging system (CrysCam UV) to facilitate the identification of protein crystals. |
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PX-04 Crystal refinement, soaking and co-crystallization |
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We provide technical supports and relevant consumables for crystal refinement, soaking and co-crystallization with small molecule compounds. |
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PX-05 Stock solutions for crystallization condition optimization |
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We provide a number of stock solutions for preparation and refinement of crystallization conditions. |
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PX-06 X-ray data collection |
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We support the shipping of pre-frozen protein crystals to the National Synchrotron Radiation Research Center, Hsinchu, for crystal testing and full X-ray data collection. |
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PX-07 Structure determination, model building, refinement and validation |
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We support crystal structure determination, model building, refinement and validation. We also support model building against cryo-EM data. |
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PX-08 Integrated services |
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The service covers all steps for protein structure determination by X-ray crystallography, ranging from recombinant protein purification, crystallization screening, crystal refinement, X-ray data collection, and structure determination, refinement and validation. |
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CS. Customization services |
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ASPC provides routine crystallization screening trials, and, on a collaborative basis, more advanced physicochemical, structural and functional analyses. (Please see the attachment for details) |
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| Package services |
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| Item # |
Item name |
Deluxe |
Basic |
SEC‐MALS only |
| CF-01 |
SDS-PAGE |
O |
O |
O |
| CF-02 |
UV scan & 260/280 |
O |
O |
O |
| CF-03 |
Tycho NT.6 |
O |
O |
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| CF-04 |
Far-UV CD spectrum |
O |
O |
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| CF-05 |
DSF Tm screen |
O |
O |
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| CF-06 |
AUC (SV mode) |
O |
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| CF-07 |
SEC |
O |
|
O |
| CF-08 |
SEC-MALS |
O |
|
O |
| Protein requirement |
| Concentration (mg/ml) |
1 |
1 |
1 |
| Volume (μL) |
1000 |
500 |
350 |
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