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An agarose gel electrophoresis method for the separation of arabinogalactan proteins
Author(s) -
Stöhr Christine,
Snogerup Lars,
Pennell Roger I.,
Kjellbom Per
Publication year - 1996
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.1996.10050943.x
Subject(s) - membrane , chromatography , isoelectric focusing , chemistry , agarose , arabinogalactan , free flow electrophoresis , electrophoresis , gel electrophoresis , polyacrylamide gel electrophoresis , biochemistry , gel electrophoresis of proteins , polysaccharide , enzyme
A method for resolving plasma membrane associated arabinogalactan proteins (AGPs) has been developed. Plasma membranes purified by aqueous polymer two‐phase partitioning were first subjected to Triton X‐114 fractionation. The resulting water phase contained all detectable plasma membrane‐bound AGPs. Plasma membrane AGPs were then resolved in an SDS‐agarose gel electrophoresis system (SDS‐AGE). For separating plasma membrane AGP species of the same apparent molecular weight but with different net charge, a two‐dimensional electrophoresis system was used, utilizing isoelectric focusing in an immobilized pH gradient in the first dimension and SDS‐AGE in the second dimension. These methods enabled the separation of individual plasma membrane AGPs. In comparison, SDS‐PAGE methods left AGPs as unresolved high molecular‐weight smears. The methods described here may help to establish some basic features of AGPs, such as the number, organization, and protein and carbohydrate characteristics of plasma membrane AGPs, as well as the relationship between plasma membrane and extracellular AGPs.