A specific stain for the detection of nonheme iron proteins in polyacrylamide gels.
Nonheme iron proteins can be visualized as blue bands in native polyacrylamide gels using a staining method that is both simple and rapid. The reaction of potassium ferricyanide with protein-bound iron atoms to form royal blue complexes occurs almost instantaneously and is sensitive enough to detect 1 microgram of analytical-grade ferritin and 2 micrograms of purified ferredoxin from cyanobacteria. No special treatment of reagents or apparatus was necessary. On comparison, this stain was found to be more specific than the Ferene S stain, not detecting bovine serum albumin even when present as a hundredfold excess over ferritin. The method was found to be effective for isoelectric focusing gels as well.
D000818:Animals; D001798:Blood Proteins / Q000032:analysis* / Q000302:isolation & purification; D002352:Carrier Proteins / Q000032:analysis* / Q000302:isolation & purification; D002417:Cattle; D002863:Chromogenic Compounds; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005288:Ferredoxins / Q000032:analysis* / Q000302:isolation & purification; D005292:Ferricyanides; D005293:Ferritins / Q000032:analysis* / Q000302:isolation & purification; D005312:Fetal Blood / Q000737:chemistry; D006736:Horses; D006801:Humans; D007501:Iron / Q000032:analysis*; D033862:Iron-Binding Proteins; D007525:Isoelectric Focusing / Q000379:methods; D008832:Microchemistry / Q000379:methods; D009211:Myoglobin / Q000032:analysis* / Q000302:isolation & purification; D012394:Rosaniline Dyes; D012710:Serum Albumin, Bovine / Q000032:analysis* / Q000302:isolation & purification; D013194:Staining and Labeling; D014168:Transferrin / Q000032:analysis* / Q000302:isolation & purification; D033863:Transferrin-Binding Proteins; D014227:Triazines
D001798:Blood Proteins; D002352:Carrier Proteins; D002863:Chromogenic Compounds; D005288:Ferredoxins; D005292:Ferricyanides; D033862:Iron-Binding Proteins; D009211:Myoglobin; D012394:Rosaniline Dyes; D014168:Transferrin; D033863:Transferrin-Binding Proteins; D014227:Triazines; D012710:Serum Albumin, Bovine; C031748:Ferene-S; D005293:Ferritins; D007501:Iron; C004692:coomassie Brilliant Blue; C028033:potassium ferricyanide