Complete extraction in a form suitable for liquid scintillation counting of tritium-labeled proteins from polyacrylamide gels.
Large (200 mm3) slices of polyacrylamide gels crosslinked with N,N'-diallyltartardiamide which contain tritium-labeled protein are readily solubilized in periodic acid for liquid scintillation counting of radioactivity, but the apparent recovery of label never exceeds 82%. Extraction of the slices with two commercial solubilizers at 60 degrees C gave recoveries of 82-90% which were not improved by prolonged incubation. Treatment of the slices at ambient temperature with 1.0 ml of 2% sodium periodate for 30 min followed by the addition of 0.7 ml of aqueous tetrabutylammonium hydroxide (40% w/v) gives solutions which can be immediately counted at 35% efficiency with low background and with 100% recovery of tritiated protein
D004591:Electrophoresis, Polyacrylamide Gel; D010504:Periodic Acid; D011506:Proteins / Q000302:isolation & purification*; D012588:Scintillation Counting; D012995:Solubility; D014316:Tritium
D011506:Proteins; D014316:Tritium; D010504:Periodic Acid