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Protein Expression & Purification


Products >  Protein_Expression_and_Purification >  Purify_Phospho-_and_Glycoproteins >  Phosphoprotein_Enrichment_Kit

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.

Each column has a maximum binding capacity of ~4 mg of phosphorylated protein.

Highly Selective Enrichment of Phosphoproteins

The Phosphoprotein Enrichment Kit may be used with any mammalian cell type. Cell lines tested include NIH 3T3, HEK 293, HeLa, Cos-7, and Jurkat. The enrichment procedure is highly efficient, as demonstrated by Western blotting analyses. Using a colorimetric phosphate detection method, we found the majority of the phosphoprotein in the eluate; negligible traces were detected in the wash fraction.

Phosphoprotein Affinity Columns yield a concentrated solution of phosphoprotein that can be analyzed by several different methods, including mass spectrometry and two-dimensional polyacrylamide gel electrophoresis (2D-PAGE).

At-A-Glance Documents Images & Data Resources

Features

  • 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

Additional Information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.


 
 
Products
Cat. # Product Package Size Price # of Units Select
635624 Phosphoprotein Enrichment Kit 6 Preps $284.00
635666 Phosphoprotein Enrichment Starter Kit 1 Purification $65.00
635626 Phosphoprotein Kit - Buffer A 500 mL $145.00
 

 

 


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