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
