Suitability of staining techniques for the detection and quantitation of nonhistone high mobility group proteins.
Three different staining techniques were compared for the detection of nonhistone high mobility group (HMG) proteins after acidic urea-polyacrylamide gel electrophoresis. Silver staining after glutaraldehyde fixation provides the highest detection sensitivity. Because of the acid solubility of HMG proteins special care has to be taken concerning fixation. Staining with colloidal CBB G-250 according to Neuhoff et al. is superior in sensitivity and reliability of quantitation when compared with noncolloidal Coomassie Brilliant Blue R-250. High detection sensitivity and reproducibility of quantitation are prerequisites for studying the tissue-specific expression of HMG proteins. In the present study tissue-specific differences in the molar amounts of various HMG proteins in thymus and erythrocytes of the chicken are documented by application of the methods tested.
Institute of Medical Chemistry and Biochemistry, University of Innsbruck, Austria.
A
A
Csordas
M
M
Pedrini
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H
Grunicke
D000818:Animals; D002645:Chickens; D003102:Colloids; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D006609:High Mobility Group Proteins / Q000032:analysis* / Q000302:isolation & purification; D006657:Histones / Q000302:isolation & purification; D012394:Rosaniline Dyes; D012834:Silver; D013194:Staining and Labeling
D003102:Colloids; D006609:High Mobility Group Proteins; D006657:Histones; D012394:Rosaniline Dyes; D012834:Silver; C024757:Coomassie brilliant blue R; C004692:coomassie Brilliant Blue