Isoelectric Focussing Assignment Help And Online Tutoring

Isoelectric Focussing

This method is used to separate protein according to their iso-electric points. A stable pH gradient is established in the gel by the addition of ampholytes. A protein mixture is placed in a well on the gel. With an applied electric field, protein enters the gel and migrates until each reaches a pH equivalent to its pI. Iso-electric focusing and SDS electrophoresis are combined sequentially in a process called two-dimensional electrophoresis. It is a more sensitive method than either of the above mentioned electrophoresis; it separates protein of identical molecular weight differing in Pi or vice-versa.

Such as an example of mixtures which is used as gel are called Ampholine and Pharmalyte have mixtures of 600 to 700 different homologues of amphoteric compounds with a spectrum of isoelectric points between 3 and 10 form a pH gradient under the influence of the electric field. Their buffering capacities is very high at their isoelectric points. Their molecular weights below 1 kDalton and these are hydrophilic.

The various techniques through which it is achieved are discussed in brief as follows:

1. Spectroscopy
2. Mass Spectrometry
3. Chromatography
4. Electrophoresis
5. Isoelectric Focussing
6. Microscopy
7. Scatchard Analysis
8. FRET
9. Blotting

Email Based Assignment Help in Analytical Biotechnology

To submit Analytical Biotechnology assignment click here