Protiens

Lecture 6 Working with proteins OBJECTIVES: At the end of these lectures, you should be subject to: mark methods of separation and purification Describe editorial chromatography Describe isoelectric focusing Explain quantification of proteins Describe analysis of proteins by SDS/ rogue Describe western blotting Why purify a protein? To claim its function To analyze its physical properties To determine its place For industrial or therapeutic applications Protein Purification Object: to divulge a especial(a) protein from all some other proteins and cell components there are many an(prenominal) proteins (over 4300 genes in E. coli) A given protein poove be 0.001-20% of summarize protein Other components: nucleic acids, carbohydrates, lipids, little(a) molecules Proteins are found in different states and locations: water-soluble, insoluble, tissue layer bound, DNA bound, in organelles, cytoplasmic, periplasmic, atomic Proteins dif fer in: Size Shape germinate Properties  go IN THE PURIFICATION OF A TYPICAL oil-soluble PROTEIN 1. Homogenization: furbish up crude extract. 2. Differential Centrifugation: Remove membranes, nuclei, greathearted organelles (mitochondria), and so forth stall free extract 3- Fractionation establish on protein size, charge, etc.

Many factors play role: pH, temperature, salt conc., and other factors 4. ammonium ion sulfate precipitation: (1) Proteins often become little soluble when the ionic strength is increased to very postgraduate levels. venturesomeness points vary by protein, so purification can be achieved. (2) Prec ipitates can be re-dissolved in a teensy vo! lume, so concentration can be achieved. (3) Dialyze the re-dissolved proteins against a baseborn salt buffer. - Dialysis is a procedure that separate proteins (large size) from small solutes 5. editorial Chromatography First you need protein from cells: Differential Centrifugation Protein purification: tower chromatography -The to the highest degree powerful method for fractionation - Solid var. (Matrix): porous...If you want to disembowel a full essay, order it on our website: BestEssayCheap.com

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