Premium
Improved rehydration procedure for polyacrylamide gels in presence of urea: Demonstration of inherited human prealbumin variants by isoelectric focusing in an immobilized pH gradient
Author(s) -
Altland Klaus,
Banzhoff Angelika,
Hackler Rolf,
Rossmann Ute
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.1150050613
Subject(s) - urea , immobilized ph gradient , chromatography , polyacrylamide , chemistry , isoelectric focusing , volume (thermodynamics) , polymerization , chemical engineering , polymer , polymer chemistry , biochemistry , organic chemistry , physics , quantum mechanics , engineering , enzyme
Polyacrylamide gels containing immobilized pH gradients (IPGs) are usually extensively washed in deionized water after polymerization and dried when longer storage is required. The original volume of the gel is readjusted by rehydration in water or a solution of appropriate composition and subsequent evaporation, if necessary, by using a fan for weight control. A more convenient procedure is to put the dried gels back into their casting mold used for polymerization and to fill the mold with the appropriate rehydrating solution. Rehydration to the original volume is completed within 30 min for gels of a thickness of 0.5 mm. With gels to be rehydrated in the presence of urea, it was found that water enters the gel matrix faster than urea; consequently, adjustment to the original gel volume at the required urea concentration is not possible in an excess of rehydration solution, but is easily achieved by the modified rehydration procedure. It was also possible to infuse a urea gradient into the gel matrix perpendicular to the pH gradient axis. The effect of urea on the patterns of inherited human prealbumin variants is demonstrated.