Final answer:
Rubisco protein can be detected in fractions after separation through PAPE, involving ammonium sulfate precipitation to reduce Rubisco content and MOAC for phosphoprotein enrichment, followed by SDS-PAGE, Pro-Q Diamond staining, and mass spectrometry.
Step-by-step explanation:
Detection of Rubisco Protein in Separated Fractions
The detection of the Rubisco protein in fractions after separation can be achieved through a prefractionation-assisted phosphoprotein enrichment (PAPE) procedure. The process includes a prefractionation step using ammonium sulfate precipitation to reduce the presence of Rubisco, followed by metal oxide affinity chromatography (MOAC) for enrichment of phosphoproteins, including those with low abundance. Common techniques such as SDS-PAGE followed by staining with Pro-Q Diamond phosphoprotein stain or mass spectrometry analysis can help identify and quantify the presence of phosphoproteins and specifically Rubisco in the separated fractions. To assess the reproducibility and efficiency of enrichment, a comparison of protein abundance is done using quantitative values based on spectral counting and confirmed with principal component analysis and scatter plots for technical reproducibility.