Pierce™ Phosphoprotein Enrichment Kit - FAQs

No. The metal and procedure used to charge the columns are proprietary.

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No. For best results, use the column only one time.

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No. Chelating agents such as EDTA and EGTA will cause metal to leach from the column and hence, reduce binding. Reducing agents such as DTT, 2-mercaptoethanol and TCEP also reduce binding to the column. Remove incompatible substances from the sample by desalting before applying to the column.

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Phosphatase and protease inhibitors are not required. If desired, add inhibitors to the Lysis/Binding/Wash Buffer as indicated in the product instructions.

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You can use RIPA buffer to lyse cells; however, dilute the lysate with the Lysis/Binding/Wash Buffer included in the kit for compatibility with the column.

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Typical yields are 10-25% of the total protein, depending on cell stimuli, cell type and binding characteristics of the specific phosphoproteins.

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Each of the 10 columns in the kit can process a single sample containing up to 4 mg of total protein. The kit includes Lysis/Binding/Wash Buffer, CHAPS, Elution Buffer and 10 Thermo Scientific Protein Concentrators (7 mL/9K) for concentrating the enriched samples.

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Yes. The Phosphoprotein Enrichment Columns bind phosphotyrosine, phosphoserine and phosphothreonine residues.

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The columns in the Thermo Scientific Phosphoprotein Enrichment Kit contain a proprietary metal that interacts with negative charges from phosphate groups. The optimized buffer conditions enable specific capture of phosphoproteins from complex biological samples.

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From 2 mg of total protein, the expected yield is 300 µg when using the Pierce Phosphoprotein Enrichment kit.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

The columns in the kit contain a proprietary metal that interacts with negative charges from phosphate groups. The optimized buffer conditions enable specific capture of phosphoproteins from complex biological samples.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Enrichment of specific proteins or protein complexes can most easily be accomplished by using target-specific immobilized antibody or PTM specific affinity binding. We offer several kits targeting phosphorylated proteins, alpha- linked mannose and terminal glucose residues, N-acetyl glucosamine, polyubiquinated proteins, ATP binding sites, and active serine hydrolase enzymes. Choose the right protein enrichment kit for your application using the following link (http://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-purification-isolation/protein-purification/protein-enrichment.html).

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.