Update profile
    Change password
    Sign out

Our content has moved! Check out our protein biology page on lifetechnologies.com  Go now >

Pierce Phosphoprotein Staining Kit

Green bands indicate specific, in-gel detection of phosphorylated proteins.

Pierce Phosphoprotein Staining Kit.

Thermo Scientific Pierce Phosphoprotein Staining Kit is for specific, colorimetric, in-gel detection of phosphoserine and phosphothreonine residues in phosphorylated proteins.

The Pierce Phosphoprotein Staining Kit is a colorimetric stain to specifically detect proteins separated by polyacrylamide gel electrophoresis (SDS-PAGE) that are phosphorylated at serine and/or threonine residues. Staining is achieved by first hydrolyzing the phosphoprotein-phosphosphoester linkage using 0.5N NaOH in the presence of calcium ions. The gel containing the newly formed insoluble calcium phosphate is then treated with ammonium molybdate in dilute nitric acid. The resulting insoluble nitrophospho-molybdate complex is finally stained with a solution of the basic dye, methyl green. The reagents in this kit hydrolyze the phosphoester linkage of phosphoserine and phosphothreonine. Phosphotyrosine is not hydrolyzed and cannot be detected with this kit. Although the staining method is not sufficiently sensitive to detect subtle changes in phosphorylation of low-abundance proteins, the kit provides a convenient system for assessing purification and phosphorylation status of phosphoproteins during preparative workflows.


  • Specific for pSer and pThr phosphoproteins – for use with SDS-PAGE to detect abundant serine- and threonine-phosphorylated proteins in the sample
  • Easy-to-follow protocol – produces results in approx. three hours, yielding phosphoproteins that are identified as green to green-blue bands
  • Qualitative staining – detection limit depends on the extent of phosphorylation and protein size and abundance; detects 40 to 80ng phosvitin and 80ng to 160ng beta-casein per band
  • Controls included – complete kit includes one positive control protein (phosvitin) and one negative control protein (soybean trypsin inhibitor) for method validation
  • Compatible with double-staining –phosphoprotein-stained gels can be secondarily stained with Thermo Scientific GelCode Blue Stain Reagent (Part No. 24590) for total protein profiling


  • Seven solutions (3L total), plus 2 vials of lyophilized protein (1mg each)


  • Evaluate the progress of a phosphoprotein purification process
  • Identify approximate molecular weights of most abundant phosphoproteins in a sample

Product Details:

Protocol summary for the Pierce Phosphoprotein Staining Kit.
Protocol summary for the Thermo Scientific Pierce Phosphoprotein Staining Kit.


Specific staining of phosphoproteins.
Specific detection of phosphorylated proteins with the Thermo Scientific Pierce Phosphoprotein Staining Kit. An identical selection of proteins was run on two gels and stained with either the Phosphoprotein Stain (left) or Thermo Scientific GelCode Blue Stain for total protein detection (right). Arrows denote lanes containing phosphoproteins. A: soybean trypsin inhibitor (negative control), B: bovine serum albumin, C: phosvitin, D: histone III-S, E: ovalbumin, F: β-casein and G: prestained protein molecular weight markers. Note that coomassie dye does not detect heavily phosphorylated proteins, such as phosvitin (right, lane C).

The limit of detection of this stain depends on several factors, including the molar amount of phosphate loaded onto the gel and the accessibility of these groups within the electrophoresed sample to hydrolysis during the staining procedure. For several proteins, the calculated lower molar limit of phosphate detection varied more than five-fold and, therefore, lower limits of detection must be determined empirically for each phosphoprotein.


  1. Simpson, R.J. (2003). Proteins and Proteomics. A Laboratory Manual. Cold Spring Harbor Laboratory Press. pp. 201-202.

Related Resources:

Protein Gel Stains Selection Guide
Protein gel electrophoresis technical guide
Protein phosphorylation technical guide

Related Products:

Phosphoprotein Enrichment Kit
Fe-NTA Phosphopeptide Enrichment Kit
TiO2 Magnetic Phosphopeptide Enrichment Kit
Phosphoprotein Phosphate Estimation Assay Kit

Ordering Information
Product # Description Pkg. Size Instructions MSDS CofA Price
24552 Pierce Phosphoprotein Control Set
Formulation: Vials of lyophilized phosvitin (40kDa phosphoprotein) and soybean trypsin inhibitor (STI, 20kDa), 1mg each
Sufficient For: Phosphoprotein control lanes in at least 10 gels

Kit Contents:
Phosvitin positive control protein, 1mg
STI negative control protein, 1mg
2-protein set Product Instructions for product #24552 Pierce Phosphoprotein Control Set MSDS for product #24552 Pierce Phosphoprotein Control Set Certificate of Analysis for product #24552 Pierce Phosphoprotein Control Set $220.00

Now available on lifetechnologies.com

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

Recommend this page

Follow Us
Email Sign-up  Email Announcements

You may also need
23235 - Micro BCA Protein Assay Kit

78503 - M-PER Mammalian Protein Extraction Reagent

89801 - Mitochondria Isolation Kit for Tissue

37536 - SuperBlock T20 TBS Blocking Buffer

37536 - SuperBlock T20 (TBS) Blocking Buffer


Antibodies  |   Molecular Biology   |   Cell Biology   |  Thermo Scientific  |   * Trademarks  |   Privacy Statement
© 2015 Thermo Fisher Scientific Inc.   |   3747 N Meridian Rd, Rockford, IL USA 61101   |   1-800-874-3723  or  815-968-0747