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You are here:Home » Molecular Biology » MB-Enzymes, Phosphatase » Protein Phosphatase 2A, Serine, Threonine (PP2A Ser/Thr)

Protein Phosphatase 2A, Serine, Threonine (PP2A Ser/Thr)

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Specifications

Type 2 protein phosphatases are serine/threonine phosphatases which can preferentially dephosphorylate the a subunit of phosphorylase kinase and are insensitive to inhibitor 2 treatment. Protein phosphatase 2A (PP2A) can be distinguished from PP2B and PP2C which require Ca2+ and Mg2+, respectively, by its partial activity in the absence of divalent cations. PP2A activity is potently inhibited by treatment with okadaic acid and shows much stronger activity toward histone phosphorylated by Protein Kinase C and casein phosphorylated by Protein Kinase A than other protein phosphatases.
Catalog #P9107-33D
Enzyme Dilution Buffer (Included)(P9107-33D1): Protein Phosphatase 2A, Serine, Threonine (PP2A Ser/Thr), Supplied as a liquid in 20mM MOPS, pH 7.4, 100mM sodium chloride, 60mM 2-mercaptoethanol, 1mM MgCl2, 1mM EGTA, 0.1mM MnCl2, 1mM DTT, 0.1mg/ml serum albumin, 10% glycerol.
ApplicationsSuitable for use in selective in vitro dephosphorylation of proteins. Other applications not tested.
Recommended DilutionsSelective in vitro dephosphorylation: Use 0.05-0.5u total of PP2A per reaction. Note: Phosphatase activity is inhibited by 10nM okadaic acid. Some preparations show extremely low levels of serine kinase activity (1-3fmole/min/mg), which can be abolished if needed by the addition of EDTA to the assay buffer.
Unit DefinitionOne Unit releases 1nmole of phosphate per min from 15uM [32P] labeled Phosphorylase A at 30°C.
Storage and StabilityAliquot to avoid repeated freezing and thawing.. Store at -20°C. Aliquots are stable for at least 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
DO NOT STORE AT -70ºC
SourceHuman red blood cells; Tested and found negative for Hepatitis B antigen, EBV and HIV antibody.
PurityPurified by a combination of DEAE, aminohexyl-agarose and gel filtration chromatography. Isolated as the heterodimer of 60kD (A) and 36kD (C) subunits.
Concentration~10u/100ul (10u of enzyme: ~5ug protein)
FormSupplied as a liquid in, 20mM MOPS, pH 7.5, 500mM sodium chloride, 60mM 2-mercaptoethanol, 1mM MgCl2, 1mM EGTA, 0.1mM MnCl2, 1mM DTT and 0.1mg/ml serum albumin, 10% glycerol.
10 Units of enzyme is ~5ug protein.
SpecificitySpecific for p-Ser and p-Thr proteins as compared to p-Tyr. Reported preference for sites in signal transduction kinases such as MAP kinases, steroid receptor, ion channels and p53 protein. This PP2A is highly active and displays characteristic properties of Type-2A phosphatases. Reconstitutes to ABC trimer with added B subunits. Extremely sensitive to okadaic acid (inhibition at < 10nM) and resistant to inhibitor-2. Phosphate, phosphoesters and fluoride are inhibitory. Sulfhydryl compounds are required to maintain activity.
Important NoteThis product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.


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