A Guide to Methods in the Biomedical Sciences by Ronald B. Corley

By Ronald B. Corley

Hundreds of thousands of equipment were constructed within the a number of biomedical disciplines, and people coated during this ebook symbolize the elemental, crucial and most generally used equipment in numerous diverse disciplines.

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Only one naturally occurring atom in proteins, hydrogen (1H), can be observed by NMR. Therefore, proteins are usually uniformly labeled with the other atoms that can be observed by NMR, 13-carbon or 15-nitrogen. ). A number of methods have been developed to determine whether a protein exists in native form or as part of a higher order polymer within cells. Non-reducing/non-denaturing gels and gradients These are often used to isolate proteins from cellular extracts in their native states. The identities of the component chains can then be identified by resolving them on denaturing and reducing/denaturing gels to determine if proteins are non-covalently or covalently associated, as described in detail earlier in this chapter.

C. Characterization of primary, secondary, tertiary and quaternary structures of proteins Protein sequencing (primary protein structure) Three different methods were initially developed for direct sequencing of proteins, These included the Sanger, Dansyl chloride, and Edman degradation techniques. All three were laborious, required huge amounts of starting material (grams or more), and also required long periods of time, often years, to complete the sequence of a short protein. For the development of one of the methods of protein sequencing and using it to solve the sequence and structure of insulin, Frederick Sanger received the first of his two Nobel Prizes in Chemistry in 1958.

Lectins can also 16 A GUIDE TO METHODS IN THE BIOMEDICAL SCIENCES be used as probe alternatives to antibodies for western blotting. Common lectins used for these studies include concanavalin A, a lectin isolated from jack beans which binds mannose residues, and ricin, a lectin from castor beans which binds galactose residues. Ricin is extremely toxic, so much so that it is considered a prime bioterrorist weapon. High-performance liquid chromatography (HPLC) HPLC is a generic term that refers to chromatography using fully automated equipment and “high performance” adsorbents for isolating and purifying compounds.

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