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Encompassing Amino Acids: full length
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: Assayed using BPS's PDE5A kit, Cat. # 60350. Assay conditions: 10 mM Tris-HCl,pH 7.4, 10 mM MgCl2, 0.1 mg/ml BSA,0.05% Tween-20, 200 µM cGMP, 2.5 kU 5'-nucleotidase, andserial dilutions of PDE5A1 at 37°C for 20 min.Quantified by 5'- nucleotidase cleaving the5' -GMP product and releasing thephosphate group which is detected byMalachite Green Reagent.
Description: Human PDE5A1 (GenBank Accession No. NM_001083), full length, with N-terminal GST-tag, MW=126 kDa, expressed in a Baculovirus infected Sf9 cell expression system.
Format: Aqueous buffer solution
Formulation: 45 mM Tris-HCl, pH 8.0, 124 mM DTT, 2.4 mM KCl, 18 mMglutathione, 3 mM DTT and 10% glycerol.
Genbank: NM_001083
Host Cell Line: Sf9 cells
Storage Stability: At least 6 months at -80°C.
Tags: N-terminal GST-tag
Uniprot: O76074
Unit Definition: One unit is defined as the amount of enzyme that will convert 1 pmole of 3', 5'-cGMP to 5'-GMP per minute at 37°C.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Filippi,S. et al., Endocrinology 148 (3), 1019-1029 (2007).
2. Bruder,S. et al., J. Biol. Chem. 281 (29), 19969-19976 (2006).Application Reference(s):
1. Characterization of the binding properties of T-773 as a PET radioligand for phosphodiesterase 10A (2015)
2. Active Site Coupling in PDE:PKA Complexes Promotes Resetting of Mammalian cAMP Signaling (2014)
3. Genetic deletion and pharmacological inhibition of phosphodiesterase 10A protects mice from diet-induced obesity and insulin resistance (2014)
4. A yeast-based chemical screen identifies a PDE inhibitor that elevates steroidogenesis in mouse Leydig cells via PDE8 and PDE4 inhibition (2013)
5. Analysis of substrate specificity and kinetics of cyclic nucleotide phosphodiesterases with N'-methylanthraniloyl-substituted purine and pyrimidine 3',5'-cyclic nucleotides by fluorescence spectrometry (2013)
6. The novel phosphodiesterase 10A inhibitor THPP-1 has antipsychotic-like effects in rat and improves cognition in rat and rhesus monkey (2013)
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