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Ultrathin‐layer isoelectric focusing in agarose gels and visualization of urinary alanine aminopeptidase with a modified agar overlay technique
Author(s) -
van Triet Adrianus J.
Publication year - 1984
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150050611
Subject(s) - agarose , isoelectric focusing , chromatography , agar , isoelectric point , polyacrylamide , aminopeptidase , agar gel , substrate (aquarium) , alanine , chemistry , materials science , biochemistry , enzyme , biology , microbiology and biotechnology , amino acid , bacteria , leucine , polymer chemistry , ecology , genetics
Ultrathin‐layer isoelectric focusing of the urinary alanine aminopeptidase in 100 μm agarose gels resulted in a better separation of the isoenzymes and proteins than in polyacrylamide gels. Although miniature isoelectric focusing over a 4 cm separation distance proved excellent for screening purposes, more details were observed in gels with a 10 cm separation distance. Optimum enzyme visualization was achieved with a modified overlay technique employing a 0.2% agar with substrate instead of the conventional 1 % agar overlay. After incubation the diluted agar overlay may be easily washed off without damaging the running gel.