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Computer‐aided standardization of two‐dimensional electrophoresis: Improvement of biophysical description of proteins in relative versus absolute coordinate systems
Author(s) -
Fosslien Egil,
Prasad Rameshwar,
Stastny Jaroslav
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.1150050208
Subject(s) - plotter , absolute scale , analytical chemistry (journal) , relative standard deviation , chemistry , approximation error , dimension (graph theory) , chromatography , mathematics , physics , statistics , combinatorics , computer science , atomic physics , computer graphics (images) , detection limit
Protein migration distances on two‐dimensional (2‐D) gels were measured in mm (absolute) and in relative internal standard units using creatine kinase carbamylation trains (x‐axis) as relative charge (CK‐C r ) scale and rat heart extract lines as relative molecular weight (M r ) scale (y‐axis). Variation in absolute protein location generally increases with increasing migration distance in both axes with 6–12 mm and 5–11 mm for selected spots in the x‐ and y‐axis respectively; variation in relative migration is significantly less, ranging from 0.9–2.5 mm and 3–4 mm in the x‐ and y‐axis respectively; data are from seven separations run in the same batch, measurements obtained from 1.6 × magnified projections of photographed gels. A computerized plotter was used to generate scaled relative charge ‐ relative molecular weight (C r ‐M r ) plots for each separation based upon absolute positions of internal protein standard spots (first dimension) and lines (second dimension). The results support the use of computer‐delineated‐relative coordinate systems (C r ‐Mr plots) for biophysical description of proteins separated by 2‐D electrophoresis.