Protein purification and centrifuge
Office Rm410;Centrifuge location: 3F Public area、Rm505、Rm506、Rm605、Rm607 (outside)、Rm705
Yan-Ping Shih +886-2-27855696,4101

The mission of Protein purification and centrifuge facility is to manage instruments for protein expression and purification. The facility provides instrument access, training course, consultation and technical development for IBC investigators. The facility currently houses several basic instruments Nanophotometer, tabletop ultracentrifuge, gradient master, bacterial culture incubators, cell disruptors, FPLC (GE) and FPLC (Bio-Rad) with autosampler and fluorescence detector. The High Performance Centrifuges (Beckman Coulter Avanti J-25/J-26XP) and Ultracentrifuges (Beckman Coulter Optima L/XL/LE), equipped with a full range of rotors, are located on each floor for common use.

1. NanoLyzer N2

Nanolyzer, high-pressure homogenizer is equipped with a hydraulic power system, which supplies pressurized hydraulic oil to a single acting intensifier pump. The intensifier pump amplifies the hydraulic pressure and transfers it to the liquid pressure of the product. Using high pressure (10-25 Kpsi) to squeeze the cells from the micro-flow channel after the rapid decompression. The pressurized product enters the diamond chamber with fixed geometrically micro-channels, causing the product to accelerate to high velocity which brings about the desired results of shear, impact, and cavitation. NanoLyzer is widely applied to particle size reduction, such as bacteria, cells, yeast, etc., to release the intracellular material.

2. Orbital Shaking Incubator

The Minstrel DT UV is an ultraviolet and visible crystal imaging and protein crystal monitoring system. It automatically images crystallization experiments and links images with crystallization conditions. This robotic instrument is a major advance over previous visible light microscopy technology because its UV technology can find crystals in complex drops and easily distinguish protein crystals from non-protein crystals (such as salt).

3. GE ÄKTA™ Protein Purification System – Explorer, purifier

ÄKTA™ Protein Purification System is an instrument for fast protein liquid chromatography (FPLC). Our system is equipped with four pumps for running four different solutions simultaneously, a full wavelength UV detector, a pH detector, a conductivity detectors and an automated fraction collector.

4. GE ÄKTA™ Protein Purification System – pure

ÄKTA™ Protein Purification System is an instrument for fast protein liquid chromatography (FPLC). Our system is equipped with four pumps for running four different solutions simultaneously, a full wavelength UV detector, a pH detector, a conductivity detectors and an automated fraction collector.

5. GE ÄKTA™ Protein Purification System – Avant

Flexible and intuitive chromatography system for fast purification of proteins, peptides, and nucleic acids from microgram to gram levels of target product. Affinity Chromatography, Gel Filtration/ Size Exclusion Chromatography, Ion Exchange Chromatography , Hydrophobic Interaction Chromatography and Reversed Phase Chromatography (RPC). ÄKTA™ avant is a preparative chromatography system with flow rates up to 25 ml/min is designed for process development, method optimization, and scale-up. Integrated fraction collector with cooling functionality protects purified samples. Automatic on –line buffer preparation using BufferPro reduces the time required for buffer blending and manual titration, increasing your productivity. UNICORN software is used to control automated processes on ÄKTA™ avant to increase productivity and efficiency. Design of Experiments (DoE), an experimental design tool, provides time and cost savings by capturing more precise information in fewer experiments. ÄKTA™ avant offers a complete solution for fast, high-quality protein separations while maintaining reliability and increasing process knowledge.

6. NGC Discover 10 chromatography system with autosampler and fluorescence detector Type: NGC Discover 10 and C96 autosampler (Bio-Rad) and RF-20Axs (Shimadzu)

The NGC Discover 10 System is designed for high-resolution separations for any application. This chromatography system may be used for method development and optimization, combining a multi-wavelength detector, automated buffer blending with buffer inlets, a column switching valve, and a sample pump for expanded condition scouting options during the purification of proteins, peptides, and nucleic acids. The NGC Discover 10 system allows automated buffer blending via the buffer blending valves and salt gradients via the buffer inlet valves. The pH valve enables pH measurement and ensures full control of your conditions, while the sample pump enables automated unattended loading of multiple large sample volumes using the sample inlet valve on up to five columns connected to the column switching valve. Fractionated samples can easily be collected using the BioFrac fraction collector. This system has applied a C96 autosampler and a fluorescence detector for high-throughput condition screening and detecting fluorescence protein.

7. Nano-photometer N60

Best in class microvolume spectroscopy with the convenience of a lightweight portable instrument with a built-in 7” inch LCD glove compatible touch screen with flexible unit control options. Built-in vortex provides sample uniformity of homogenous samples for the highest degree of accuracy with the smallest sample volume (1 ~ 2μl) with as little as 0.3 μl. With a variety of detection methods, including single, multi, full wavelength detection, absorption ratio, kinetics, nucleic acid quantitative: Built-in determination of dsDNA, ssDNA, RNA, oligonucleotide and other method parameters, without further setting. Protein Quantification: Built-in UV280 parameters, no need to be set separately. Xenon lamp source without warm-up. Turns on and measures immediately, with Vortex built-in to ensure sample uniformity. Full scan capability range from 200-900 nm for rapid and complete sample analysis in as little as 3.5 seconds. Exclusive sample suppression technology, the sample solution is limited to above the sample tank, do not have to worry about the impact of evaporation, more viscous or volatile samples can be used. Linux kernel software interface, support for Windows, Android and iOS operating system. No re-calibration, no need to calibrate the optical path, ensure the optical path is always fixed, no need to calibrate the instrument for life, IMPLEN guarantee lifetime accuracy.

8. High Performance Centrifuges (BECKMAN COULTER Avanti J-25/ Avanti J-26XP serials)
Following Rotors: JS-13.1, JA-14, JA-20, JA-25.15, JA-25.50, JLA-10.500, JLA-16.250, JLA-8.1000, F10BCI-6x500y FIBERLite. High-speed centrifuge is used for sample preliminary purification, which used for large sample volume, and less purity. Select the appropriate rotor and centrifuge tubes, so you can get better experimental results. The load in a laboratory centrifuge must be balanced carefully. This imbalanced force the spindle and may result in damage to the centrifuge or personal injury.
9. Ultracentrifuges (BECKMAN COULTER Optima L / XL / LE serials)

Following Rotors: SW 28 Ti, SW 41 Ti, Type 45 Ti, Type 50.4 Ti, Type 55.2 Ti, NVT™ 65 Rotor Assy, Type 70.1 Ti.

  • Fixed Angle Rotor (designated Type)—-are general-purpose rotors that are mainly used for pelleting subcellular particles and in short-column banding of viruses and subcellular organelles.
  • Vertical-Tube rotor (designated V)—-are used for pelleting, separation of density and sedimentation coefficient.
  • Swinging-Bucket Rotor (designated SW)—-are hold tubes parallel to the axis of rotation, used for isopycnic and, in some cases, rate zonal separations when run time reduction is important.
  • Near-Vertical Rotor (designated NV)—-are designed for gradient centrifugation when there are components in a sample mixture that do not participate in the gradient.

Selection of a rotor depends on a variety of conditions, such as sample volumn, number of sample components to get separated, particle size, desired run time, desired quality of separation, type of separation, and the centrifuge in use.

10. Bentch-top ultracentrifuge    Location: Rm413

The Optima MAX-TL can be used in applications including separation of subcellular particles, viruses and viral particles, rate-zonal separation of proteins in sucrose gradient, separation of lipoproteins, pelleting of RNA through a CsCl gradient, isopycnic separation of plasmid DNA and others. With a maximum speed of 120,000 rpm and maximum rcf of 657,000 x g, the instruments control speed within ±50 rpm of set point. Temperatures can be set from 0° to 40°C in 1°C increments, and controlled within ±2°C of set point (after equilibrium).

11. Gradient Master & Swinging bucket rotors (SW55Ti)

The Gradient Master™ forms 6 identical linear density gradients in 1-2 min using a technique called tilted tube rotation. The gradient forming process involves layering the two end point solutions (i.e. 5% and 30% sucrose) directly in the centrifuge tube, the tube is capped to lock in the solutions, placed in a holder and rotated at a high tilt angle for a fixed time and speed. This process is easy, fast and highly reproducible. There is no competing device with this capability.