go to the first exercise
Lectures can cover the theoretical aspects of protein purification and laboratory classes can teach the practical techniques, but there are other topics which are difficult to teach by conventional methods. In order to purify any protein you need to know which separation techniques are likely to be most effective under the circumstances and, probably more important, which techniques are not. This knowledge cannot be picked up by following a fixed recipe for a class practical. It requires some thought and usually comes with experience, generally during postgraduate research.
The material presented here has been developed in the School of Biochemistry and Molecular Biology at the University of Leeds. It aims to guide students through a simulation of some of the more commonly-used protein separation techniques and to let them experiment with the simulation. It starts off by letting the user examine how a simple mixture of proteins behaves during gel filtration and ion-exchange chromatography and then goes on to allow the design and testing of full purification protocols using more complex mixtures of proteins.
It is assumed that students are familiar with the theoretical background to the most common separation techniques, enzyme assays etc. and that they understand the concept of the isoelectric point of proteins. Bear in mind that this is a laboratory simulation. IT IS NOT A SELF-CONTAINED TUTORIAL ON PROTEIN PURIFICATION.The exercises rely on a Java applet and its associated files. You must use a Java-enabled browser such as Netscape or Internet Explorer versions 4 or later.
The applet has been written to use the Swing user interface classes. Experience has shown that the Java Virtual Machines packaged with browsers can be unreliable. For this reason, the applet is designed to use the Sun Java Plug-in, which you can download from http://java.sun.com/products/plugin/index-1.4.html, which will ensure that you are using a reliable version of Java.
Make sure that you have the Java Plug-in installed and then click here to start the applet in a new window.
The exercises are illustrated with images depicting the program running on a Windows computer, but if you are running it on a different computer - don't worry, it operates in an identical manner.
If you have started the applet, you are ready to work through the exercises.
These pages are best viewed with Netscape or Internet Explorer, version 4.0 or later, maximized, using a screen resolution of 1024 x 768. There are a lot of images, so if you have a slow connection or if you are using a dial-up connection, you might be better off downloading the pages with an application such as HTTP Weazel so that you can view them off-line. If you do this, remember that if the pages are updated, your copy will become out of date.
Contact the author if you want details or if you have any comments on the material presented here.Here is some information about designing your own mixtures.