Kinase Activity Profiling Services

Are you looking for thorough solutions for your questions regarding the role of kinases in your research? Measuring kinase activity provides deeper understanding of cellular signaling and target interaction.

Mechanistic insights

Kinases are the most intensively studied protein targets and are the basis of numerous types of therapies. However, the traditional approaches study the abundance of proteins rather than their activity. This results in a knowledge gap on how cell signaling really works and a partial understanding of your drug.

Building on 15 years of experience, PamGene has developed a unique technology with unprecedented sensitivity, making real-time measurement and understanding of kinase activity possible. The multiplex properties of our chip enable us to look at the bigger picture of cellular signaling. It makes you see the activity changes in pathways, and allows the generation of a kinase-based response biomarker.

As fellow scientists we are eager to learn about your challenges. We’re happy to schedule an online meeting to see how our approach can benefit your research.

Applications

Thanks to our unique and proven technology we will provide you with robust measurements, which can be applied in numerous of applications bringing your research and insights to the next level.

Kinase inhibitor selectivity profiling
Pathway elucidation
Drug-target interaction
Target discovery and identification
Biochemical identification
Biomarker discovery and identification

Our workflow

We offer Contract Research Services tailored to your needs and research questions.

Intake meeting – with our scientists to define your research goals
Experimental design development – based on our extensive experience and fitted for your goals
Collecting samples – by the client following our easy-to-use sample preparation protocols
Process and analysis – using our unique and proven PamStation12 technique
Results meeting – data will be shared and discussed with you by our scientists

“While quantitative measurements of transcripts and proteins are key to investigate the state of a biological system, they do not inform the “active” state of regulatory networks.” “Recent advancements of high-throughput protein kinase activity profiling platforms allow for a broad assessment of protein kinase networks in complex biological systems. In conjuncation with sophisticated computational modeling techniques, these profiling platforms provide datasets that inform the active state of regulatory systems in disease models and highlight potential drug targets.”

Learn more about this recently published review paper in ‘The current opinion in pharmacology’

Read further

Mechanically activated Piezo1 channels of cardiac fibroblasts stimulate p38 mitogen-activated protein kinase activity and interleukin-6 secretion.

Blythe NM, Muraki K, Ludlow MJ et. al.
J Biol Chem. Oct 4. pii: jbc.RA119.009167. doi: 10.1074/jbc.RA119.009167. EPub. (2019)

Publication

Profiling Activity of Cellular Kinases in Migrating T-Cells.

Chirumamilla, C. S. et al.
Methods Mol. Biol. 1930, 99–113 (2019).

Publication

Inactive immune pathways in triple negative breast cancers that showed resistance to neoadjuvant chemotherapy as inferred from kinase activity profiles.

Sawada T, Hilhorst R, Rangarajan S et. al.
Oncotarget. Sep 28;9(76):34229-34239. (2018)

Publication

The benefits our technology brings

Kinase activity-based – Direct inhibitor effects on kinases can be measured
Sensitive – Only small amounts of protein input (0.5 to 5 μg per array) are required, thus making the assay more sensitive than alternative approaches e.g. MS-based phosphoproteomics.
Unbiased – Unlike antibody-based approaches, prior knowledge of involved kinases and targets is not required as we cover a total of 340 phosphosites.
No specific antibodies – The quality of the data is not dependent on the specificity of phospho-antibodies.
Full-length kinases – We measure kinase activity of full-length proteins from lysates of several cell lines and tissues. This is not offered by other recombinant-based kinase activity assays.

Our technology

Kinases are the most intensively studied protein targets. We have developed a unique method to understand them. Our sensitive platform uses lysates obtained from only a few thousand cells or small amounts (2ug) of animal, xenografts or human tissues and cells to simultaneously determine the activity of all kinases present in these lysates.

This is accomplished by incubating the lysates across 196 to 144 tyrosine or serine/threonine kinase peptide substrates immobilized on the PamChip® microarray platform. Our 3D microarrays are spotted with peptides (i.e. phosphosites) that represent kinase targets. Kinases present in the lysates will phosphorylate the peptide substrates which are detected using fluorescently labelled antibodies. The phosphosites on the PamChips are human-derived and there is significant overlap with other organisms.

Based on current online knowledge, we compiled a comprehensive, integrated database (DB) of potential kinases that are linked to the peptides on the PamChips. This corresponds to ~350 unique kinases in literature, covering the majority of the kinome.

Kinase activity profiling cases we’ve worked on

We have over 15 years of experience in the field of kinase science and contributed to over 100+ publications with top European, North American and Japanese Institutes.

To learn how our services contributed to different studies, request access to our case report.

Case report

Mechanically activated Piezo1 channels of cardiac fibroblasts stimulate p38 mitogen-activated protein kinase activity and interleukin-6 secretion.

Blythe NM, Muraki K, Ludlow MJ, Stylianidis V, Gilbert HTJ, Evans EL, Cuthbertson K, Foster R, Swift J, Li J, Drinkhill MJ, van Nieuwenhoven FA, Porter KE, Beech DJ, Turner NA.
J Biol Chem. Oct 4. pii: jbc.RA119.009167. doi: 10.1074/jbc.RA119.009167. EPub. (2019)

Publication

Profiling Activity of Cellular Kinases in Migrating T-Cells.

Chirumamilla, C. S. et al.
Methods Mol. Biol. 1930, 99–113 (2019).

Publication

Inactive immune pathways in triple negative breast cancers that showed resistance to neoadjuvant chemotherapy as inferred from kinase activity profiles.

Sawada T, Hilhorst R, Rangarajan S, Yoshida M, Tanabe Y, Tamura K, Kinoshita T, Shimoyama T, van Beuningen R, Ruijtenbeek R, Tsuda H, Koizumi F.
Oncotarget. Sep 28;9(76):34229-34239. (2018)

Publication

Our workflow

Intake meeting – with our scientists to define your research goals
Experimental design development – based on our extensive experience and fitted for your goals
Collecting samples – by the client following our easy-to-use sample preparation protocols
Process and analysis – using our unique and proven PamStation12 technique
Results meeting – data will be shared and discussed with you by our scientists