In Vitro Phosphorylation Assays

In Vitro Phosphorylation Assays

In Vitro Phosphorylation Assays


Creative BioMart provides a comprehensive portfolio of assays for in vitro analysis of phosphorylation in both eukaryotic and prokaryotic organisms to help elucidate dynamic phosphorylation events. The advanced technology platform and our scientific team’s unparalleled expertise in kinase/phosphatase biology and pharmaceutical research enable us to help our customers achieve scientific breakthroughs.


Protein phosphorylation is the most common and well-studied post translational modification (PTM). It is estimated that about 30% of the proteome is affected by phorphorylation. The phosphoryl group from ATP is transferred to the amino acid side chain by a kinase and removed by a phosphatase. Kinases participate in elaborate phosphorylation cascades by acting on a number of different substrates and undergoing mutual phosphorylation and activation among different types of kinases. Although kinases have been shown to be involved in signal transduction and other complex regulatory networks, our global understanding of protein phosphorylation has until recently been extremely limited.

Phosphorylation plays a key role in both homeostasis and disease by regulating protein function, altering its structural conformation, and modulating the nature and strength of protein-protein interactions. Phosphorylation at different residues can lead to different consequences. Because of this, quality tools for assessing kinase activity and protein phosphorylation are essential. So far, each technique that has been developed performs well in different applications. However, how to choose the method that is best suited for the experimental design is an issue worthy of attention, which often depends on project objectives, expertise, access to instruments, and budget.

In vitro phosphorylation assay showing cross-phosphorylation among kinase.

Figure 1. In vitro phosphorylation assay showing cross-phosphorylation among kinase. (Shi L, et al., 2014)

Service Offering

Creative BioMart is dedicated to providing our customers with end-to-end services for in vitro phosphorylation analysis to help them save time and effort. Our experienced scientists will work closely with our customers to create the perfect plan for their project and accelerate their journey to success. If you can’t find the assay format of your interest, please fill out the online inquiry form and let us know more details.

Assay Formats


Western Blot

  • Popular and gold standard for measuring phosphorylation levels qualitatively.
  • Allow for the identification of specific phosphorylation sites.

Enzyme-Linked Immunosorbent Assay (ELISA)

  • High specificity, high sensitivity, high throughput, and quantifiable.
  • Simple data interpretation.

Chemiluminescence Immunoassay

  • Wide dynamic range and high degree of automation.
  • High signal intensity and high specificity.

Flow Cytometry

  • Rapid, quantitative, and single cell analysis.
  • Multiplex detection and high throughput.

Immunocytochemistry (ICC/IHC)

  • Assessment of multiple proteins within a cell or tissue.
  • Both fluorescent and colorimetric detection techniques can be used.

Antibody Arrays

  • Multiplex detection and high throughput.
  • Without the requirement of special equipment.

Mass Spectrometry (MS)

  • Large dynamic range and high sensitivity.
  • Comprehensively evaluation of phosphorylation in complex biological samples independent of antibodies.

Creative BioMart is an advanced biotech company within the field of kinase/phosphatase biology. We offer our expertise in establishing assays for phosphorylation analysis. The extensive experience of our scientific team translates into the robust wealth of knowledge that we use to provide you with the highest quality of service. We also provide custom phosphorylated antibody development, antibody pairing and assay validation services. If you are interested in any of our services or have additional questions, please do not hesitate to contact us.


  • Shi L, et al. Cross-phosphorylation of bacterial serine/threonine and tyrosine protein kinases on key regulatory residues. Frontiers in microbiology, 2014, 5: 495.
  • Ptacek J, et al. Global analysis of protein phosphorylation in yeast. Nature, 2005, 438(7068): 679-684.
For research use only. Not intended for any clinical use.