Kozulić M, Kozulić B, Mosbach K
Institute of Biotechnology, ETH Hönggerberg, HPT, Zurich, Switzerland.
Anal Biochem. 1987 Jun;163(2):506-12. doi: 10.1016/0003-2697(87)90255-7.
We describe in detail the synthesis of an acrylic monomer, N-acryloyl-tris(hydroxy-methyl)aminomethane (NAT), which was successfully used for the preparation of gels for electrophoresis and isoelectric focusing. The polymerization kinetics and transparency of the poly(NAT) gels crosslinked by N,N'-methylenebisacrylamide (Bis) are also shown. Poly(NAT)-Bis gradient (4-24%) gel resolves proteins according to their size. The exclusion limit of this gel is slightly over 3 X 10(6), which is more than threefold higher than the exclusion limit of the polyacrylamide gradient gel of the same concentration. The gel made of 6% NAT and 3% Bis represents a suitable matrix for isoelectric focusing. These results demonstrate that poly(NAT)-Bis gels could be advantageously used in those applications where the extensive sieving by the polyacrylamide matrix is not desir desirable.
我们详细描述了一种丙烯酸单体N-丙烯酰基-三(羟甲基)氨基甲烷(NAT)的合成方法,该单体已成功用于制备电泳和等电聚焦凝胶。文中还展示了由N,N'-亚甲基双丙烯酰胺(Bis)交联的聚(NAT)凝胶的聚合动力学和透明度。聚(NAT)-Bis梯度(4-24%)凝胶可根据蛋白质大小分离蛋白质。这种凝胶的排阻极限略高于3×10⁶,比相同浓度的聚丙烯酰胺梯度凝胶的排阻极限高三倍多。由6%NAT和3%Bis制成的凝胶是等电聚焦的合适基质。这些结果表明,聚(NAT)-Bis凝胶可有利地用于那些不希望聚丙烯酰胺基质进行广泛筛分的应用中。