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Thermo Scientific™ Pierce™ Phosphoprotein Enrichment Kit

Catalog No. PI90003
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10 Column Kit
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PI90003 10 Column Kit
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Catalog No. PI90003 Supplier Thermo Scientific™ Supplier No. 90003

Description

Purify (enrich) phosphorylated proteins from mammalian cell and tissue lysates using phospho-specific IMAC affinity resin cartidges or kit.

Perform efficient enrichment of phosphorylated proteins derived from mammalian cells and tissues with Thermo Scientific™ Pierce Phosphoprotein Enrichment Kit.

The kit is an effective system for isolating the phosphoprotein fraction of mammalian cell and tissue lysates. The included lysis-binding buffer and proprietary affinity resin produce superior yields with negligible non-specific binding. The kit uses a convenient spin column format, and the resulting phosphorylated protein sample is compatible with downstream applications, including Western blotting, mass spectrometry (MS), 2D-PAGE, and protein arrays. The affinity resin is also available as pre-packed chromatography cartridges for use with automated liquid chromatography (LC) systems.

Highlights:

  • Specific – low nonspecific protein contamination from complex biological samples, such as cell culture lysate and mouse tissue extract

  • Fast – easy spin format enables enrichment of phosphorylated proteins in less than 2 hours

  • Superior yield – achieves higher yields than other commercially available phosphoprotein isolation kits

  • Convenient format – complete kit offering pre-dispensed spin columns, buffers, reagents and ultrafiltration columns for enrichment

  • Compatible – downstream applications include mass spectrometry, Western blotting and 2D-PAGE

Specifications

Content And Storage Upon receipt store at 4°C.
Product Line Pierce
Quantity 10 Column Kit
Target Phosphoprotein
Type Phosphoprotein Enrichment Kit
Stationary Phase Proprietary Ligand
Sufficient For 10 Reactions
Format Spin Column
Column Type Affinity, Agarose Resin

Frequently Asked Questions (FAQs)

Can I recharge the column in the Thermo Scientific Phosphoprotein Enrichement kit?

No. The metal and procedure used to charge the columns are proprietary.
Can I reuse or recharge the columns when using the Thermo Scientific Phosphoprotein Enrichment kit?

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

Are samples containing EDTA or reducing agents compatible with the Thermo Scientific Phosphoprotein Enrichment kit?

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.

Do I need phosphatase inhibitors when using the Thermo Scientific Phosphoprotein Enrichment kit?

Phosphatase and protease inhibitors are not required. If desired, add inhibitors to the Lysis/Binding/Wash Buffer as indicated in the product instructions.
What other type of lysis buffer can I use with the Thermo Scientific Phosphoprotein Enrichment kit?

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.

What type of yield should I expect with the Thermo Scientific Phosphoprotein Enrichment kit?

Typical yields are 10-25% of the total protein, depending on cell stimuli, cell type and binding characteristics of the specific phosphoproteins.
How many samples can I process and what is included in the Thermo Scientific Phosphoprotein Enrichment kit?

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.
Will the columns bind phosphoproteins modified on tyrosine residues?

Yes. The Phosphoprotein Enrichment Columns bind phosphotyrosine, phosphoserine and phosphothreonine residues.
How does the Thermo Scientific Phosphoprotein Enrichment kit work?

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.

What is the expected phosphoprotein yield when using the Pierce Phosphoprotein Enrichment kit?

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

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.
Can I isolate my proteins based on their post-translational modification (PTM) or active binding site?

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).

For Research Use Only. Not for use in diagnostic procedures.