By Nathan P. Kaplan, Nathan P. Colowick, William B. Jakoby, Meir Wilchek

ISBN-10: 0121818977

ISBN-13: 9780121818975

The significantly acclaimed laboratory average, Methods in Enzymology, is likely one of the such a lot hugely revered courses within the box of biochemistry. seeing that 1955, each one quantity has been eagerly awaited, often consulted, and praised by way of researchers and reviewers alike. The sequence includes a lot fabric nonetheless suitable at the present time - really a vital book for researchers in all fields of lifestyles sciences

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Additional info for Affinity Techniques - Enzyme Purification: Part B

Sample text

Otherwise, small clumps of gel remain which are difficult to disperse even with prolonged stirring. ,3 The term "functional group density" (or "group density") will be used to express the degree of substitution of amide groups in terms of millimoles of introduced group per dry gram of original polyacrylamide. The term "specific capacity, ''1 or simply "capacity," has often been used to mean the same thing. However, it would seem preferable to reserve "capacity" as an expression of the maximum amount of a moiety that the adsorbent is able to bind in its eventual application.

O C O N H R ~OH The reactive species formed in the activation step have different degrees of stability. After acid treatment a product is formed that can be freeze-dried and stored for long periods of time. Agarose (and crosslinked agarose) activated and thus stabilized is commercially available (Pharmacia). A number of variations differing slightly in detail have been published and are claimed to represent improvements. Under certain conditions such as high concentration of cyanogen bromide, a high degree of substitution may be reached.

Darkly colored material should be purified as follows: Dissolve 25 g of the crude anhydride in 125 ml of benzene. 5 g of activated charcoal and stir for 1 hour at room temperature. Filter and reduce the volume of filtrate to about 60 ml by boiling (in a hood). Cool the solution to 20 ° and add gradually, with stirring, 180 ml of ethyl ether. Cover and allow to stand until crystallization starts. Place the well-covered container in a refrigerator for 1 hour. Collect crystals by suction filtration, wash with about 30 ml of ether and dry under vacuum.

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Affinity Techniques - Enzyme Purification: Part B by Nathan P. Kaplan, Nathan P. Colowick, William B. Jakoby, Meir Wilchek


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