By Nathan P. Kaplan, Nathan P. Colowick, William B. Jakoby, Meir Wilchek
The significantly acclaimed laboratory normal, Methods in Enzymology, is among the so much hugely revered guides within the box of biochemistry. when you consider that 1955, each one quantity has been eagerly awaited, often consulted, and praised via researchers and reviewers alike. The sequence includes a lot fabric nonetheless correct this present day - really a vital book for researchers in all fields of existence sciences
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Extra info for Affinity Techniques - Enzyme Purification: Part B
The beads are offered in various size ranges and porosities as described in their catalog. Type P-2, for example, is highly cross-linked and, when fully swollen, has a molecular weight exclusion limit of 1800; higher P values (P-4 through P-300) have progressively fewer cross-links and higher molecular weight exclusion limits. 32 COUPLING REACTIONS AND GENERAL METHODOLOGY  macromolecules can be separated effectively on polyacrylamide adsorbents. Because of the softness of low cross-linked beads and consequent resistance to flow in columns, one should select bead porosities no greater than that needed to obtain an effective interactive surface.
Preparation of adsorbents containing low molecular weight ligands may be exemplified by sulfanilamide-Sepharose 6B. ~7 Coupling. 0, at 50 °, containing 500 mg of sulfanilamide. The mixture is gently stirred at 50 ° for 2 hours. 0 containing 1 M NaCl, and 500 ml of 1 M NaC1 at 50 °. The coupled sulfanilamide is 550 /zmoles per gram of dry gel according to the nitrogen content as determined by the Kjeldahl method. 3 M sodium carbonate at pH 10 containing 200 mg of protein. The suspension is gently stirred with a magnetic stirrer at room temperature for 2 hours.
The polyacrylamide gel beads are purchased under the trade name, Bio-Gel P, from Bio-Rad Laboratories, 32nd and Griffin Avenue, Richmond, California 94804 and their distributors. The beads are offered in various size ranges and porosities as described in their catalog. Type P-2, for example, is highly cross-linked and, when fully swollen, has a molecular weight exclusion limit of 1800; higher P values (P-4 through P-300) have progressively fewer cross-links and higher molecular weight exclusion limits.
Affinity Techniques - Enzyme Purification: Part B by Nathan P. Kaplan, Nathan P. Colowick, William B. Jakoby, Meir Wilchek