Hottest acrylamide gel electrophoresis

Acrylamide gel electrophoresis

polyacrylamide gel electrophoresis is an electrophoretic method using polyacrylamide gel as the supporting medium. On this support medium, the separation can be based on the molecular size and molecular charge of the separated substance

polyacrylamide gel can't get accurate experimental results, which has the following advantages:

① polyacrylamide gel is a macromolecule polymerized by acrylamide and N, n 'methylenebisacrylamide. The lattice of gel is a carbon carbon polymer with amide side chain, and there is no or little ionic side group, so the electroosmosis effect is relatively small and it is not easy to interact with the sample

② as polyacrylamide gel is a kind of synthetic substance, the concentration ratio of monomer can be adjusted before polymerization to form a cross chain structure to varying degrees, and its porosity can change in a wide range. According to the size of the molecule of the substance to be separated, the appropriate composition of gel can be selected, so that it not only has a suitable porosity, but also has better mechanical properties than the double tube structure of the drop weight impact tester. Generally speaking, it contains acrylamide The mechanical properties of 5% gel are suitable for separating substances with a molecular weight range of less than 10000 to 1million. For proteins with a molecular weight of less than 10000, gel containing acrylamide 1 is used to further enhance industrial upgrading%, while gel containing acrylamide 4% can be used for proteins with a particularly large molecular weight. The macroporous gel is fragile, and the small pore gel is difficult to take out from the tube. Therefore, when the concentration of acrylamide increases, the double acrylamide can be reduced to improve the mechanical properties of gel

③ polyacrylamide is stable to heat in a certain concentration range. Gel is colorless and transparent, easy to observe, and can be directly determined by detector

④ acrylamide is a relatively pure compound, which can be refined to reduce pollution. The raw materials for synthesizing polyacrylamide gel are acrylamide and methylenebisacrylamide. Acrylamide is called monomer, and methylenebisacrylamide is called crosslinking agent. In aqueous solution, monomer and crosslinking agent form gel through polymerization initiated by free radicals

in the reaction process of polyacrylamide gel formation, a catalyst is required. The catalyst includes initiator and another accelerator. Initiators provide initial free radicals in the formation of gel. Through the transmission of free radicals, acrylamide in the downstream of the petrochemical industry becomes free radicals to initiate polymerization. Accelerators can accelerate the release of free radicals from initiators. The compatibility of commonly used initiators and accelerators is shown in the table below: