Phosphatase Assays

Reaction Biology offers phosphatase assays for compound screening and profiling for drug discovery. Our phosphatase assays are based on the dephosphorylation of the fluorinated MUP (methylumbelliferyl phosphate) derivative which serves as a substrate for most protein phosphatases. Dephosphorylation of MUP generates the fluorescent DiFMUP (6,8-difluoro-4-methylumbelliferyl phosphate) which is quantified for the determination of phosphatase activity.

Protein phosphatases play important roles in cell signaling processes in tandem with kinases. Different than kinases, phosphatases were long considered undruggable because their active site is highly conserved resulting in selectivity issues and an unfavorable molecular composition leading to bioavailability issues. The discovery of an allosteric SHP2 inhibitor paved the road for the creation of further allosteric inhibitors of phosphatases. Our activity assays are an essential tool for phosphatase profiling for allosteric inhibitor discovery.

  • Fluorescence-based phosphatase activity assays
  • High-throughput compatible phosphatase screening
  • Custom-tailored phosphatase assay development possible
  • Deliverable: % inhibition (single point) or IC50 value determination

At Reaction Biology, we offer phosphatase profiling and screening services for pharma, biotech, and academic institutions with a commitment to quality data and quality service. Our global business development team is available to discuss your phosphatase screening needs with you today.

List of Phosphatase Assays (13)

Target HGNC Symbol Additional Synonyms Data
DUSP22/MKPX DUSP22 Dual specificity protein phosphatase 22, JNK-stimulatory phosphatase-1, Low molecular weight dual specificity phosphatase 2, Mitogen-activated protein kinase phosphatase x, JSP1, LMWDSP2 Data sheet
PP1A PPP1CA Serine/threonine-protein phosphatase PP1-alpha catalytic subunit, PPP1A Data sheet
PP1B PPP1CB Serine/threonine-protein phosphatase PP1-beta catalytic subunit Data sheet
PP2A alpha/PPP2R1A Complex PPP2CA/PPP2R1A Complex PP2A alpha: Serine/threonine-protein phosphatase 2A catalytic subunit alpha isoform; PPP2R1A: Serine/threonine-protein phosphatase 2A 65 kDa regulatory subunit A alpha isoform Data sheet
PTPN1/PTP1B PTPN1 Tyrosine-protein phosphatase non-receptor type 1, Protein-tyrosine phosphatase 1B Data sheet
PTPN11/SHP2 (E76K)-FL PTPN11 (E76K) Tyrosine-protein phosphatase non-receptor type 11, Protein-tyrosine phosphatase 1D, Protein-tyrosine phosphatase 2C, SH-PTP2, SH-PTP3, PTP2C, SHPTP2 Data sheet
PTPN11/SHP2-CD PTPN11 Tyrosine-protein phosphatase non-receptor type 11, Protein-tyrosine phosphatase 1D, Protein-tyrosine phosphatase 2C, SH-PTP2, SH-PTP3, PTP2C, SHPTP2 Data sheet
PTPN11/SHP2-FL PTPN11 Tyrosine-protein phosphatase non-receptor type 11, Protein-tyrosine phosphatase 1D, Protein-tyrosine phosphatase 2C, SH-PTP2, SH-PTP3, PTP2C, SHPTP2 Data sheet
PTPN12/PTP-PEST PTPN12 Tyrosine-protein phosphatase non-receptor type 12, Protein-tyrosine phosphatase G1, PTN12 Data sheet
PTPN2/TC-PTP PTPN2 Tyrosine-protein phosphatase non-receptor type 2, T-cell protein-tyrosine phosphatase, PTN2, PTPT Data sheet
PTPN6/SHP1 PTPN6 Tyrosine-protein phosphatase non-receptor type 6, Hematopoietic cell protein-tyrosine phosphatase Protein-tyrosine phosphatase 1C, Protein-tyrosine phosphatase SHP-1, HCP, PTP1C Data sheet
PTPN7/LC-PTP PTPN7 Tyrosine-protein phosphatase non-receptor type 7, Hematopoietic protein-tyrosine phosphatase, PTN7 Data sheet
PTPRC/CD45 PTPRC Receptor-type tyrosine-protein phosphatase C, Leukocyte common antigen, T200 Data sheet

Phosphatase Assay Details

Assay Format

Principle of phosphatase assay with the fluorogenic substrate difmup.

The fluorinated MUP derivative is used as a substrate for a large range of protein phosphatase assays. The reaction product of DiFMUP is fluorescent after dephosphorylation.

Assay Setup

Setups: Single-dose screening in duplicates or IC50 value determination with 5 or 10 concentrations. Other screening formats are available upon request.

Controls: No inhibitor (DMSO vehicle) control and for every assay, one target-specific control compound is tested in 10-dose IC50 format. 

Turnaround time: 10 business days for standard projects. Expedited scheduling and data delivery can be arranged prior to the commencement of the studies.

Report: The raw data, % enzyme activity and control compound IC50 values will be reported in Excel format for single-dose assays. For IC50 orders, raw data, IC50 values, and curve fitting will be delivered in Excel format. Assay conditions, target, and substrate information are available upon request. Requirements for this information should be noted prior to the commencement of the study.

Screening facility: This assay is performed at our screening facility in Malvern, PA, US.

Compound requirements: In brief, for a standard project, 20 µl of a 10 mM DMSO stock or solid material is needed. Less material is needed for large scale screening. Please refer to our FAQs for information regarding compound preparation and shipping.