PDE1C Assay Kit

CAT: 0421-60312Size: 96 ReactionsDry Ice: YesHazardous: No
CAT#:0421-60312Size:96 Reactions
Selected
PRICE
AVAILABILITY: InStock
24/48H Stock Items & 2 to 6 Weeks non Stock Items.
Product image 1
1 / 1
Background
Phosphodiesterases (PDEs) play an important role in the dynamic regulation of cAMP and cGMP signaling. PDE1C is a calmodulin-dependent PDE that is expressed principally in human myocardium. Phosphodiesterases catalyze the hydrolysis of the phosphodiester bond in dye-labeled cyclic monophosphates. Beads selectively bind the phosphate group in the nucleotide product. This increases the size of the nucleotide relative to unreacted cyclic monophosphate. In the polarization assay, dye molecules with absorption transition vectors parallel to the linearly-polarized excitation light are selectively excited. Dyes attached to the rapidly-rotating cyclic monophosphates will obtain random orientations and emit light with low polarization. Dyes attached to the slowly-rotating nucleotide-bead complexes will not have time to reorient and therefore will emit highly polarized light.
Description
The PDE1C Assay Kit is a fluorescence polarization (FP), homogeneous, 96-well assay kit designed for the screening and profiling of PDE1C (Phosphodiesterase 1C) inhibitors. This assay takes advantage of a specific fluorescent phosphate-binding nanoparticle. The kit contains enough purified recombinant PDE1C, fluorescent probe, PDE assay buffer, Binding Agent, and diluent for 100 reactions.Figure 1: Illustrationof the PDE1C Assay Kit principle.The assay uses a fluorescein-labeled cyclic adenosine monophosphate (cAMP-FAM for PDE1C), in which the phosphate group is engaged within the cyclic nucleotide. This is a very small molecule that rotates fast (low FP) . PDE1C catalyzes the hydrolysis of the phosphodiester bond in the cyclic nucleotide and frees the phosphate group. In a second step the free phosphate group is recognized by a specific phosphate-binding nanobead (Binding Agent) leading to the formation of a large complex, with restricted movement (high FP) . FP is proportional to PDE1C activity.This assay requires a fluorescent microplate reader capable of measuring fluorescence polarization (FP) and equipped with the required parts to read the FP signal. For more information FP technology, visit our Tech Note:FP, assay principles and applications.Need us to run inhibitor screens or profile your compounds against PDE1C? Check out ourPhosphodiesterase Screening Services.Note:As of December 2024, this protocol has been re-optimized for performance. Previous versions of this kit are available upon request.
UniProt
Q14123
Applications
Useful for studying enzyme kinetics and screening small molecular inhibitors for drug discovery and HTS applications.
Format
Catalog #NameAmountStorage60013PDE1C, GST-Tag*˃1 µg-80°C60200FAM-Cyclic-3′, 5′-AMP**1.2 nmoles**-80°C60393PDE Assay Buffer (Incomplete) 25 ml-20°C60390PDE Binding Agent100 µl4°C60391PDE Binding Agent Diluent (cAMP) 10 ml4°C827350.5 M DTT200 µl-20°C79685Low binding, black 96-well plate1Room Temp.* The concentration of protein is lot-specific and will be indicated on the tube containing the protein.**FAM-Cyclic-3′, 5′-AMP is provided as a powder. The vial will need to be resuspended in 600 µl of Complete PDE Assay Buffer before use.
Shipping Conditions
-80°C
Storage Conditions
At least 6 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.
Contraindications
The final concentration of DMSO in the assay should not exceed 1%.Fluorescent compounds that have λex=470 nm and detection at λem=528 nm can interfere with the readouts.It is recommended that the compound alone is tested to determine any potential interference of the compound on the assay results.

Related Products

CatalogName
61312PDE1C (Mouse) Assay Kit
60705PDE1C TR-FRET Assay Kit
60384-1PDE4C Assay Kit
60311PDE1B Assay Kit
60384-2PDE4C Assay Kit
79501-1PDE6C Assay Kit
79501-2PDE6C Assay Kit
60384PDE4C Assay Kit
79501PDE6C Assay Kit
60310PDE1A Assay Kit

Alternative Products

CatalogName
GWB-PS7A7FPDE1C Assay Kit
137330PDE1C Assay Kit, BioAssayTM
LKU607342Phosphodiesterase 1C, Calmodulin Dependent (PDE1C) Magnetic Luminex Assay Kit
MBS2160216-01Phosphodiesterase 1C, Calmodulin Dependent (PDE1C) Magnetic Fluorescence Assay Kit
MBS8562080-01PDE1C
MBS8501463-01PDE1C Antibody
MBS3236322-01PDE1C Peptide
512601PDE1C discontinued
MBS515626-01PDE1C, Active
571740-01PDE1C (HRP)

Popular Products