NCSB

Overview

There are four main laboratory facilities at the NCSB.  They are X-ray, NMR, Protein Expression and Purification (PrEP) and Crystal Screening and Growth (CSG).

X-ray Structure Solving

The NCSB provides two state of the art x-ray facilities.  One is located at the Eppley Institute which is part of the University of Nebraska Medical Center in Omaha and the other is located at the University of Nebraska at Lincoln. Click on the link(s) below for the available equipment and other details.

Eppley X-ray Laboratory

UNL X-ray Laboratory

NMR Structure Solving

The NMR Laboratory is based in the Eppley Institute.

NMR Laboratory

Protein Expression and Purification (PrEP)

The NCSB PrEP lab is located primarily in the Lied Transplant Center.  It has the necessary instruments for expressing large quantities of recombinant protein ( fermentors, shakers, centrifuges and liquid chromatography systems ). Specialized instruments are also used in the Durham Research Center (analytic ultra-centrifuge).

Key Equipment

	New Brunswick ScientificInnova Shakers The Innova Shakers can handle up to 8 4l flasks each and have temperature control systems that allow from 4C to 80C at up to 400 RPMs. Typically they are configured to provide a range of sample sizes from test tubes to 4l flasks.
	New Brunswick Scientific BioFlo 4500 Fermentor The BioFlo fermentor can grow cultures up to 20l. The system can self autoclave. Besides being able to self autoclave it can automatically adjust the pH, oxygen level, and temperature. It is used when there is a need for large scale expression.
	Sorvall RC 5C plus Flow Through Centrifuge The Sorvall centrifuge has a flow through attachment that allows cultures grown by the fermentor to spun down into pellets as they are drained. The centrifuge has a number of rotors that allow for conventional pelleting of flask grown cultures as well.
	Akta explorer HPLC The Akta explorer HPLC allows for complex protein purification schemes to be implemented automatically. The lab has a wide range of columns that contain most of the commonly used resins. Additionally, we have to super loops (40ml and 10ml) that allow for faster loading and processing of larger samples.
	Akta FPLC The FPLC allows for much simpler purification schemes to be implemented the HPLC. All of the columns and super loops used by the HPLC are available for the FPLC

Crystal Screening and Growth (CSG)

The NCSB CSG lab is located in the Lied Transplant Center.  It has the instruments necessary for determining and refinement of crystal conditions.  Some key pieces of equipment include the Art Robbin's Phoenix crystallization robot, several large crystal incubators and a high resolution polarized imaging microsope.

Key Equipment

	Wyatt Technologies Dynamic Light Scattering Instrument The DLS can determine the relative populations in a solution.  It is useful for optimizing protein solutions for crystallization by making it possible to identify conditions where there is only one species in solution.  These cases have a much higher probability of crystallization.  The DLS can also determine the rough size of the population.
	Wyatt Technologies Size Exclusion Chromatography and Multi-Angle Light Scattering Instrument The SECMALs can determine the various species in solution and also measure very accurate molecular weights for those species.  This information can be used to determine the composition of complexes in solution at biologically relevant concentrations and the individual complexes can be separated with the fraction collector and used in crystallization experiments. It can autoclave itself
	Art Robbins Instruments Phoenix The Phoenix is used to automatically setup 96 well formatted trays.  Generally we use 200nl drops but the system can dispense down to 20nl drops.  Using 200nl drops allows five times more crystallization experiments to be setup with the same amount of solution compared to normal approaches.
	Formulatrix RockImager 54 The Formulatrix RockImager automatically images crystallization experiments that use SBS formated plates and stores the results in a relational database with the RockMaker front end. RockMaker stores all of the experimental information related to a plate so that this information can be used when combined with the imaging data to develop random screen experiments that will improve the probability of crystallization.