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FAQs for Protein Tools Technical Reference for Protein Tools Enzyme Finder NEBcutter NEBuffer Chart Double Digest Finder PCR Selection Tool Isoschizomers DNA Sequences and Maps REBASE Protein Phosphatase 1 (PP1)

Home > Products > Protein Tools > Protein Phosphatases/Inhibitors > Protein Phosphatase Inhibitor 2 (I-2) Protein Phosphatase Inhibitor 2 (I-2) Catalog # Size Concentration Price Qty   P0755L 250 μg 1 mg/ml $448.00 P0755S 50 μg 1 mg/ml $112.00 Prices are in US dollars and valid only for US orders. Download: MSDS PDF Type 1 protein phosphatase (PP1) specific Inhibitor Rabbit skeletal muscle, recombinant (E. coli) Supplied with 10X Reaction Buffer Description: Protein Phosphatase Inhibitor-2 (I-2) specifically and instantaneously inhibits the catalytic subunit of type I protein phosphatase (PP1) at nanomolar concentrations. In contrast, the inhibition of native forms of PP1, type 1 holoenzymes, is time dependent and requires much more I-2. I-2 inhibits PP1 from mammals, yeast and flies in an identical manner and can be used for identification of type I protein phosphatases from many different species (1-3). I-2 is a 204 amino acid heat stable protein. Recombinant I-2 has identical inhibitory properties as native rabbit muscle I-2 (2). I-2 is functionally tested in the inhibition assays of PP1 (NEB #P0754). Source: Isolated from a strain of E. coli that carries the coding sequence for rabbit skeletal muscle I-2 polypeptide under the control of T7 expression system (kindly provided by Dr. A. A. DePaoli-Roach) (3). Properties & Usage Molecular Weight: Theoretical: 22,800 daltons and Apparent: 31,000 daltons Specific Activity: 1,000,000 units/mg Reaction & Storage Conditions Reaction Conditions: 1X PP1 Reaction Buffer Supplemented with 1 mM MnCl2 and 5 mM Caffeine Incubate at 30°C. 1X PP1 Reaction Buffer: 50 mM HEPES 100 mM NaCl 2 mM Dithiothreitol 0.1 mM EGTA 0.025 % Tween-20 pH 7.5 @ 25°C Concentration: 1 mg/ml Storage Conditions: 50 mM HEPES 0.1 mM EGTA 50% Glycerol 0.01% Brij 35 pH 7.0 @ 25°C Storage Temperature: -20°C Notes General notes: Supplement with 1 mM MnCl2 and 5 mM caffeine (required only if phosphorylase a is the substrate). I-2 has been purified to > 95% homogeneity as determined by SDS-PAGE and Coomassie Blue staining. Recommended long term storage -70°C; avoid repeated freeze/thaw cycles. Usage notes: PP1 or cell extracts should be diluted so that the concentration of phosphatase activity is less than 0.1 U/ml in the assays. One unit is the amount of PP1 that hydrolyzes 1 nmol of substrate in 1 minute. The diluted extracts must be preincubated with I-2 for 10-15 minutes prior to assay in order to achieve complete inhibition (1). Quality Control for Current Lot Quality control values for a specific lot can be found on the datacard which accompanies each product. Protease Activity: After incubation of 5 µg of I-2 with a standardized mixture of proteins for 2 hours at 30°C, no proteolytic activity could be detected by SDS-PAGE. Phosphatase Activity: After incubation of 5 µg of I-2 with 50 mM p-Nitrophenyl Phosphate (NEB #P0757) for 2 hours at 30°C, no phosphatase activity could be detected by spectrophotometric analysis. References Cohen, P. (1991) Methods in Enzymology, 201, 389-398. Park, I., et al. (1994) J. Biol. Chem, 269, 944-954. Connor, J.H., et al. (2000) J. Biol. Chem, 275, 18670-18675. Companion Products Protein Phosphatase 1 (PP1) New England Biolabs, Inc. is an ISO 9001 and ISO 14001 Certified Company. NEB certifies that it is a small business in accordance with the US Small Business Administration and 13 CFR 121.201