Alphascreen cgmp phosphodiesterase assay using alphascreen cgmp supplement with anti-cgmp antibody

AlphaScreen cGMP Phosphodiesterase AssayUsing AlphaScreen cGMP Supplement with Anti-cGMP Antibody TION NOTE
Figure 1. Principle of the AlphaScreen cGMP PDE assay. The cGMP PDE enzyme depletesthe unlabeled cGMP competitor, allowing the two AlphaScreen beads to bind via the anti-cGMP antibody interaction. As a result, an intense AlphaScreen signal is generated. Introduction
second form is of cytosolic nature2.
w w w. p e r k i n e l m e r. c o m
* Please note that for compound screening purposes, we recommend adding 0.5 µLof test compounds to 4.5 µL of cGMP Materials and methods
reaction Buffer. If the test compounds are Results and discussion
Figure 2. Titration of enzyme and substrate in the PDE assay using a detection time of 2 hours. confirming the stability of the assay.
Figure 3. Time Course of PDE enzymatic activity. Left panel: AlphaScreen Signal (cps); Right Panel: amount of cGMP hydrolyzed (pmoles) Figure 4. Inhibition Curves for PDE V w w w. p e r k i n e l m e r. c o m
1) Ghalayini IF, ATP-regulated
module (ARM) of the atrial
natriuretic factor receptor
guanylate cyclase, Int J Impot
Nitric oxide-cyclic GMP path-
way with some emphasis on
cavernosal contractility,
Peptides 2005 Jun;26(6):969-84.
A Simple Statistical Parameter
for Use in Evaluation and
Validation of High Throughput
Screening Assays,
J Biomol Screen Apr 1999; 4: 67-73.
Corbin JD, Inhibition of Cyclic
GMP-Binding Cyclic GMP-
Specific Phosphodiesterase
(Type 5) by Sildenafil and
Related Compounds
, Mol
1999 Jul; 56(1):124-30.
Cyclic GMP Phosphodiesterase
Inhibitors. 2. Requirement of
6-Substitution of Quinazoline
Derivatives for Potent and
Selective Inhibitory Activity
Figure 5. Screening validation of AlphaScreen cGMP PDE assay. Representative Z’ graph for a 384 manual assay. PerkinElmer Life and
Analytical Sciences
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