SDD-AGE

The SDD-AGE (of English. Semi-denaturing detergent agarose gel electrophoresis , agarose gel electrophoresis halbdenaturierende detergenzvermittelte ') is a biochemical method in which SDS-resistant protein complexes such. B. in amyloidoses or prion diseases , separated by gel electrophoresis in an agarose gel and detected.

principle

SDS-resistant protein complexes are assemblies of several proteins which do not denature at an SDS concentration of 2% (m / V) at room temperature . Their molar masses are mostly between 200 kilodaltons and 4000 kilodaltons, which is why an agarose gel with relatively large pore diameters is used. Following the electrophoresis, analogous to the Western blot with a polyacrylamide gel , a protein transfer to a PVDF membrane and an immunostaining of the proteins on the membrane take place. This enables conformational variants of the proteins of a protein aggregate to be differentiated.

Individual evidence

1. ↑ Dmitry S. Kryndushkin, Ilya M. Alexandrov, Michael D. Ter-Avanesyan, Vitaly V. Kushnirov: Yeast [PSI +] prion aggregates are formed by small Sup35 polymers fragmented by Hsp104. In: The Journal of Biological Chemistry. Vol. 278, 2003, ISSN  0021-9258 , pp. 49636-49643, PMID 14507919 , doi : 10.1074 / jbc.M307996200 .
2. ↑ Randal Halfmann, Susan Lindquist : Screening for amyloid aggregation by Semi-Denaturing Detergent-Agarose Gel Electrophoresis. In: The Journal of Visualized Experiments. Vol. 17, 2008, ISSN  1940-087X , p. 838, PMID 19066511 ; PMC 2723713 (free full text), doi : 10.3791 / 838 .