Phosphoprotein enrichment & purification kit

The Phosphoprotein Enrichment Kit provides an effective affinity-based procedure for isolating phosphorylated proteins from mammalian cells and tissues. Each kit includes a complete set of buffers along with six high-capacity columns for enrichment of both cytosolic and membrane-bound phosphoproteins, regardless of the amino acid modified—including serine, tyrosine, or threonine.

Our enrichment procedure offers a number of advantages:

• The procedure is fast; the average cell-to-sample purification time is less than 2 hours.
• It is also straightforward, consisting of four main steps: adding Extraction/Loading Buffer to the cell or tissue pellet to extract total cellular protein, loading the extract on an affinity column, washing, and finally eluting the bound phosphoprotein with a detergent-free Elution Buffer.
• A single buffer—Extraction/Loading Buffer—is used for both the protein-extraction and affinity-column steps, making buffer exchange unnecessary. This saves time and prevents sample loss.
• The procedure is nondenaturing, so phosphoproteins remain folded throughout the process, even during the extraction and elution steps.

Overview

• Nondenaturing protocol maintains protein conformation and solubility
• Ideal for use in many downstream applications, including mass spectrometry and 2D-PAGE
• Increase the sensitivity of your phosphodetection experiments

Applications

• Mass spectrometry sample preparation
• Phosphorylation analysis
• 2D-PAGE sample preparation