PDE6C Human Phosphodiesterase Enzymatic LeadHunter Assay - TW

Item: 156130

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OVERVIEW

DOCUMENTS/REFERENCES

Biological Information

Background Information:

The phototransduction cascade of retinal rods is a particularly accessible G protein signaling system. Upon absorption of a photon, photoexcited rhodopsin activates many transducin molecules which activate PDE6 to remove its inhibitory constraint on the α and β catalytic subunits of PDE6. Hydrolysis of cGMP by PDE6 lowers the cytoplasmic cGMP concentration and closes cGMP-gated cationic channels in the rod plasma membrane, which reduces the inward current.

Family:

PDE

Sub Family:

Phosphodiesterase

Class:

Cyclic Nucleotide Signaling

Protein Name:

Phosphodiesterase 6C

Uniprot Number:

P51160

Protein Aliases:

cGMP phosphodiesterase 6C

Gene Name:

PDE6C

Gene ID:

5146

Gene Aliases:

PDEA2; ACHM5; COD4

Accession Number:

NM_006204

Organism:

Human

Construct Details:

Full length, Recombinant

Assay Information

Assay Type:

Biochemical

Assay Sub Type:

Enzymatic

Functional Mode:

Antagonist

Detection Method:

Fluorescence Polarization

Platform:

Plate reader

Measured Response:

Testing Information

Procedure Summary:

Human recombinant PDE6C expressed in insect Sf9 cells is used. The test compound and/or vehicle is preincubated with 75 ng/ml enzyme in Tris-HCl buffer pH 7.2 for 15 minutes at 25°C. The reaction is initiated by addition of 100 nM fluorescein-labeled cGMP for another 30-minute incubation period and terminated by addition of an IMAP binding solution. IMAP complexes with phosphate groups on nucleotide monophosphate generated from cyclic nucleotides through PDE activity. Determination of the amount of complex formed is read spectrofluorimetrically at 470 nm/525 nm. Compounds are screened at 10 µM.

Substrate:

0.1 µM FAM-cGMP

Incubation:

30 min at 25°C

Control Inhibitor:

Zaprinast

Criteria For Significance:

≥ 50% of max stimulation or inhibition

Test Concentration / Dose:

Suggested concentration for initial testing is 10 uM

Test Sample Requirements:

Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).

Minimum Order Quantity:

Turnaround Time

Standard:

15 Business Days

Reference Compound Data

PubChem ID

135399235

Name

Zaprinast

Measurement

0.21μM - IC50

Clinical Relevance

Therapeutic Area:

CNS/Addiction/Pain

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei