PP2Calpha Human Phosphatase Enzymatic LeadHunter Assay - TW
OVERVIEW
DOCUMENTS/REFERENCES
Biological Information
Background Information:
Protein phosphatase 2C-alpha (PP2Cα, PPM1A or PPPM1A protein phosphatase 1A magnesium-dependent alpha isoform, or MGC9201) is a Mn2+- or Mg2+-dependent protein serine/threonine phosphatase that is essential for regulating cellular stress response, gene expression and replication in eukaryotes. This phosphatase dephosphorylates, and negatively refulates the activities of, MAP kinases and MAP kinase kinases. It has been shown to inhibit activation of p38 and JNK kinase cascades induced by environmental stress. This phosphatase can also dephosphorylate cyclin-dependent kinases, and thus may be involved in cell cycle control. Overexpression of this phosphate is reported to activate the expression of tumor suppressor gene TP53/p53, which leads to G2/M cell cycle arrest and apoptosis.
Family:
Phosphatase
Class:
Protein Ser/Thr
Protein Name:
PP2Calpha
Uniprot Number:
P35813
Protein Aliases:
phosphatase 2C alpha|protein phosphatase 2C, alpha isoform
Gene Name:
PPM1A
Gene ID:
5494
Gene Aliases:
MGC9201|PP2Calpha|PP2CA
Accession Number:
NM_021003
Organism:
Human
Construct Details:
Full Length, Recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Spectrofluorimetry
Measured Response:
Fluorescence
Testing Information
Procedure Summary:
Human recombinant protein phosphatase PP2Cα expressed in E. coli is used. Test compound and/or vehicle is preincubated with 1.25 μg/ml enzyme‡ for 15 minutes at 37°C in modified Tris-HCl buffer pH 8.5. The reaction is initiated by addition of 100 μM DiFMUP for 60 minutes. Determination of the amount of DiFMU formed is read spectrofluorimetrically at 358 nm/450 nm. Compounds are screened at 10 μM.
Substrate:
100 µM DiFMUP
Incubation:
60 min at 37°C
Control Inhibitor:
(NH₄)₆Mo₇O₂₄
Criteria For Significance:
≥ 50% of max stimulation or inhibition
Test Concentration / Dose:
Suggested concentration for initial testing is 10 uM
Test Sample Requirements:
Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).
Minimum Order Quantity:
2
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Clinical Relevance
Adverse / Beneficial Action:
Inhibition of PPM1A may facilitate inflammation and cell proliferation.
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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