Ultraviolet scanning densitometry for detection, quantitation, and preparative elution of protein bands from unstained gels.
Ultraviolet scanning of gel rods was used to identify and quantify protein bands in a nondestructive manner with good precision and sensitivity. This same technique, applied on a preparative scale, allowed quantitative protein elution, by reversed electrophoresis, from gel slices completely sealed in a dialysis bag. Protein recovery approached the theoretical yield (93.5 +/- 5%), with practically no interfering substances, and the entire preparative process (first electrophoresis, densitometric scanning, and reversed electrophoresis) could be performed in approximately 6 h. Its application to human growth hormone has shown no alteration in the biological activity of this protein.
Bartolini
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P
Department of Application of Nuclear Techniques in Biological Sciences, Cidade Universitaria, São Paulo, Brazil.
Arkaten
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R
Ribela
M T
MT
D000818:Animals; D003720:Densitometry / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D013006:Growth Hormone / Q000032:analysis / Q000302:isolation & purification; D006801:Humans; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification; D051381:Rats; D012710:Serum Albumin, Bovine / Q000032:analysis; D013025:Glycine max; D014361:Trypsin Inhibitors / Q000032:analysis / Q000302:isolation & purification; D014466:Ultraviolet Rays
D011506:Proteins; D014361:Trypsin Inhibitors; D012710:Serum Albumin, Bovine; D013006:Growth Hormone