Molecular Biology Problem Solver 37. Sách được nhiều nhà khoa học có uy tín, nhiều kinh nghiệm trong nghiên cứu thực nghiệm trình bày những vấn đề thường hay phát sinh trong phòng thí nghiệm. Do vậy mà sách không trình bày các protocol hay quy trình như các sách khác, thay vào đó các tác giả sẽ trình bày các vấn đề nhằm giúp giúp đọc giả: Tự nâng cao khả năng chẩn đoán nguyên nhân khi gặp các vấn để về kỹ thuật, quy trình, hóa chất, thuốc thử trong quá trình thực nghiệm trong phòng thí. | Many non-ionic or zwitterionic detergents can be used for IEF or native PAGE to keep proteins soluble. CHAPS 3- 3-cholamidopropyl dimethylammonio -1-propanesulfonate is most often used as it is a very good solubilizer and is nondenaturing. It should be used from up to . Another very effective solubilizer is SB 3-10 decyldimethylammoniopropane-sulfonate but it is denaturing Rabilloud et al. 1997 . Other detergents designed especially for IEF on IPG gels have recently been designed and used successfully Chevallet et al. 1998 Molloy 2000 . The minimum detergent concentration for effective solubilization must be determined for each sample Rabilloud et al. 1999 . Again to learn what detergent might be effective for your sample we suggest a literature search. What Other Additives Can Be Used to Enhance Protein Solubility Some proteins are very difficult to solublize for electrophoresis. Urea can be used from 2 to 8M or . Thiourea can be used at up to 2M it greatly enhances solubility but cannot be used at higher concentration. This is because above 2 M the urea thiourea or detergent may precipitate out Molloy 2000 . The total urea concentration urea thiourea cannot be above approximately if thiourea is used with a bis gel due to these solubility constraints. AGAROSE ELECTROPHORESIS What Is Agarose Agarose an extract of seaweed is a polymer of galactose. The polymer is 1 3-linked beta -D-galactopyranose and 1 4-linked 3 6-anhydro- alpha -L-galactopyranose. The primary applications are electrophoresis of nucleic acids electrophoresis of very large proteins and immunoelectrophoresis. What Is Electroendosmosis -Mr or EEO -Mr is a measure of the amount of electroendosmosis that occurs during electrophoresis with a particular grade of agarose. Electroendoosmosis is the mass movement of water toward the cathode against the movement of the macromolecules which is usually toward the anode. High -Mr means high electroendosmosis. The mass flow of water toward the .
