Address: 45-1 Ramsey Road, Shirley, NY 11967, USA    USA: 1-631-559-9269  1-631-448-7888   Fax: 1-631-938-8127  Europe: 44-207-048-3343   Email: info@creative-biomart.com
Home > Products > Assay Kits > Activity Kits > Cyclic Nucleotide Phosphodiesterase assay Kit

Cyclic Nucleotide Phosphodiesterase assay Kit

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Cat.No.:
Kit-0667
Product Overview:
Unique, non-radioactive, colorimetric assay.Used to screen inhibitors and modulators of cyclic nucleotide phosphodiesterase activity.HTS format permitting rapid assays of large numbers of samples.
Size:
96 wells
Description:
A BIOMOL GREEN Quantizyme Assay System. The Cyclic Nucleotide Phosphodiesterase (PDE) Assay Kit combines a special dual enzyme system with the BIOMOL GREEN Reagent for phosphate detection to create a unique, non-radioactive, colorimetric assay. This HTS-friendly, mix and read system may be used to screen inhibitors and modulators of cyclic nucleotide phosphodiesterase activity. 96-well microplate format permits rapid assays of large numbers of samples.The basis for the assay is the cleavage of cAMP or cGMP by a cyclic nucleotide phosphodiesterase. The 5"-nucleotide released is further cleaved into the nucleoside and phosphate by the enzyme 5"-nucleotidase. The phosphate released due to enzymatic cleavage is quantified using BIOMOL GREEN reagent in a modified Malachite Green assay. The kit includes Type I cyclic AMP phosphodiesterase (PDE) for validation purposes.
Applications:
Colorimetric detection, HTS
Storage:
-80°C
Kit Components:
PDE Enzyme (from bovine brain) (5 x 4 U; lyophilized solid, 4 U per vial. 1 U = 1 nmol 3",5-cAMP to 5"-AMP per minute under the conditions of the linearity assay (30°C, pH 7.4, 200µM 3",5"-cAMP Storage: -70°C, avoid freeze/thaw cycles.5"-Nucleotidase (1ml; 5 kU/µl in 10mM TRIS-hydrochloric acid, pH 7.4, 125mM sodium chloride, 10% glycerol, 1mM magnesium chloride. One U will release 1 pmol phosphate per minute from 200µM 5"-AMP, 30°C in.a reaction buffer of 10mM TRIS-hydrochloric acid, pH 7.4, 0.2mM magnesium chloride). Storage: -70°C3",5"-cAMP Substrate (2ml; 1mM in assay buffer (10mM TRIS-hydrochloric acid, pH 7.4). Storage: -20 or -70°C3",5"-cGMP Substrate (2ml; 1mM in assay buffer (10mM TRIS-hydrochloric acid, pH 7.4). Storage: -20 or -70°CPDE Assay Buffer (40ml (2 x 20ml); 10mM TRIS-hydrochloric acid, pH 7.4) Storage: -70°CBiomol Green Reagent (20ml; liquid in screw-cap plastic bottle). Storage: 4°C, long-term at -70°C5"-AMP Standard (1.0ml; 100µM in 10mM TRIS-hydrochloric acid, pH 7.4 buffer). Storage: -20 or -70°C5"-GMP Standard (1.0ml; 100µM in 10mM TRIS-hydrochloric acid, pH 7.4 buffer). Storage: -20 or -70°CInhibitor (IBMX) (200µl; 200µM in assay buffer (10mM TRIS-hydrochloric acid, pH 7.4)). Storage: -20 or -70°CDesalting column and resin (1 column and 1 g of resin). Storage: room temperature, after rehydration store at 4°C½ volume microplate. Storage: Room temperature


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Optional requirements on this protein +Expand
C-fusion   N-fusion  Non-tagged
His   GST  Fc  Others
<1.0 eu/μg   <0.1 eu/μg  <0.01 eu/μg  Not required
Monomer Isolation   Dimer Isolation   Not required
>80% by SDS-PAGE   >90% by SDS-PAGE  >95% by SDS-PAGE  Others
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45-1 Ramsey Road, Shirley, NY 11967, USA
USA: 1-631-559-9269  1-631-448-7888
Europe: 44-207-048-3343
FAX: 1-631-938-8127
EMAIL: info@creative-biomart.com

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