| 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. |
| Applications | Suitable for use in selective in vitro dephosphorylation of proteins. Other applications not tested. |
| Recommended Dilutions | Selective 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 Definition | One Unit releases 1nmole of phosphate per min from 15uM [32P] labeled Phosphorylase A at 30°C. |
| Storage and Stability | Aliquot 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 |
| Source | Human red blood cells; Tested and found negative for Hepatitis B antigen, EBV and HIV antibody. |
| Purity | Purified 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) |
| Form | Supplied 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. |
| Specificity | Specific 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 Note | This 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. |
