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The voltage across wide pH range immobilized pH gradient gels and its modulation through the addition of carrier ampholytes
Author(s) -
Fawcett John S.,
Chrambach Andreas
Publication year - 1986
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.1150070605
Subject(s) - immobilized ph gradient , chemistry , ionic strength , swelling , chromatography , electrode , electrophoresis , analytical chemistry (journal) , tube (container) , materials science , composite material , biochemistry , isoelectric focusing , aqueous solution , enzyme
In pH 4–10 immobilized pH gradient (IPG) electrophoresis, one observes (a) partial failure of the sample to enter the gel; (b) which halt protein migration; (c) absence of defined transient state band patterns; (d) lateral zone spreading and distorted zone profiles; (d) uneven voltage profiles with a distinct voltage maximum; (f) difficulty in measuring pH profiles with the contact electrode. These problems are remedied through the addition of 0.1 to 1 % carrier ampholytes (CA) to the IPG gel. Problems (a) to (c) disappear in IPG tube gels. IPG in presence of CAs (ICAPG) of p I ‐range 3.5–9.5 or 6–8 yields a pH gradient, a protein pattern and a pH gradient stability essentially the same as that in IPG gels of the same pH‐range 4–10. ICAPG is insensitive to ionic strength of the sample up to 0.1 M. Gel swelling and water exudation increase in ICAPG which make ICAPG in tube gels inapplicable when CA concentrations exceed 0.1 %.