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

Molecular Biology

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

The significantly acclaimed laboratory regular, Methods in Enzymology, is without doubt one of the so much hugely revered courses within the box of biochemistry. considering 1955, each one quantity has been eagerly awaited, often consulted, and praised via researchers and reviewers alike. The sequence comprises a lot fabric nonetheless correct this present day - really a necessary ebook for researchers in all fields of existence sciences

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Chromatogr. 86, 53 (1973). 31p is the percentage of matrix substance ( w / v ) in swollen gel and is equal to 4 and 6 for Sepharose 4B and 6B, respectively. 24 COUPLINGREACTIONSAND GENERAL METHODOLOGY [2] Although most applications of cyanogen bromide coupling have involved agarose and cross-linked agarose, other hydroxyl-containing polymers may also be converted to biospecific adsorbents by coupling of suitable ligands in the same manner. , two carbohydrates. O/C -,/O~. O/)~ /O~. Carbohydrate 2) The carbohydrates may be monosaccharides or polysaccharides.

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. 34 C O U P L I N GREACTIONS AND GENERAL METHODOLOGY [3] swell to a settled or packed bed volume that ranges from 4 ml to 40 ml or so depending on the degree of cross-linkage.

The gradient substance competes with the adsorbed substances for the adsorption sites. Matrix Selection Proper selection of a matrix or carrier for the ligands is of decisive importance for the successful application of stereospecific adsorption. The ideal matrix should meet the following demands: 1. 2. 3. 4. 5. Insolubility Sufficient permeability High rigidity and suitable particle form Zero adsorption capacity Chemical stability under the conditions required for adsorption, desorption, and regeneration 6.

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