PA protein was first purified using a Centri-Spin 10 column to remove all free phosphates (Princeton Separations, Adelphia, NJ). Samples were analyzed in duplicate for cGMP-hydrolytic activity using a commercially available colorimetric cyclic nucleotide phosphodiesterase assay kit (Enzo, Farmingdale, NY) with and without 100 nM of sildenafil (Sigma-Aldrich) as previously described [6 (link), 23 (link)]. Protein concentration was determined using the Bradford method [22 (link)]. Results are shown as the PDE5-specific pmol cGMP hydrolyzed/minute/mg of total purified protein.
Colorimetric cyclic nucleotide phosphodiesterase assay kit
Manufactured by Enzo Life Sciences
The Colorimetric cyclic nucleotide phosphodiesterase assay kit is a laboratory tool used to measure the activity of cyclic nucleotide phosphodiesterase enzymes. It provides a colorimetric-based method for quantifying the hydrolysis of cyclic nucleotides.
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Lab products found in correlation
3 protocols using colorimetric cyclic nucleotide phosphodiesterase assay kit
1
Quantitative Analysis of PDE5 Activity
2
Measuring cGMP-Hydrolytic Activity of PA Protein
3
Measuring cGMP-Hydrolytic Activity of PA Protein
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PA protein was assayed for cGMP-hydrolytic activity using a commercially available kit (Enzo, Farmingdale, NY) as previously described (25 (link)). Briefly, PA protein was purified over a Centri-Spin 10 column to remove any free phosphates (Princeton Separations, Adelphia, NJ). Purified protein concentrations were determined using the Bradford method (44 (link)). Samples were assayed in duplicate for cGMP-hydrolytic activity using a commercially available colorimetric cyclic nucleotide phosphodiesterase assay kit (Enzo, Farmingdale, NY) with and without sildenafil (100nM, Sigma-Aldrich, St. Louis, MO). Results are shown as PDE5-specific pmol cGMP hydrolyzed/min/mg total protein.
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