Complete extraction in a form suitable for liquid scintillation counting of tritium-labeled proteins from polyacrylamide gels
✍ Scribed by Clive Meredith; Martin K. Johnson
- Publisher
- Elsevier Science
- Year
- 1987
- Tongue
- English
- Weight
- 344 KB
- Volume
- 162
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
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