[26] Detecting radiolabeled lipid-modified proteins in polyacrylamide gels

摘要: Publisher Summary This chapter discusses the methods for labeling cells and detecting tritium-labeled proteins after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE. The are specifically designed use with fatty acid-modified proteins, most equally applicable to other lipid modifications. Tritium-labeled precursors generally best metabolic of owing higher specific activity (relative 14 C derivatives) lower level interconversion. length period can be varied minimize Thus, fate exogenously added labeled acids is checked by analysis protein-bound label. usually done high-performance liquid chromatography (HPLC) or thin-layer (TLC) released acid hydrolysis methyl esters produced alkaline methanolysis. If comparing two myristate palmitate, a useful quick check method specificity process samples total cell lysates on SDS-polyacrylamide gels followed fluorography.