Biological Information

Background Information:

Nav1.5 is the predominant voltage-gated sodium channel expressed in cardiomyocytes and is encoded by the SCN5A gene. Structurally, Nav1.5 consists of a pore-forming α-subunit containing four homologous domains with six transmembrane segments each. It mediates the rapid inward sodium current responsible for depolarization of the cardiac action potential, playing a critical role in cardiac excitability and impulse conduction. Pathogenic variants in SCN5A can either increase or decrease Nav1.5 channel activity, disrupting cardiac electrical conduction and giving rise to a spectrum of inherited arrhythmia syndromes, including Long QT Syndrome, Brugada syndrome, sick sinus syndrome, and cardiac conduction disease. Due to its essential role in cardiac electrophysiology and safety pharmacology, Nav1.5 is a key target in cardiovascular risk assessment and drug discovery.

Family:

Ion Channel

Sub Family:

Sodium

Class:

Voltage-Gated

Protein Name:

Nav1.5

Uniprot Number:

Q14524

Protein Aliases:

Sodium channel protein type V subunit alpha; Voltage-gated sodium channel subunit alpha Nav1.5

Gene Name:

SCN5A

Gene ID:

6331

Organism:

Human

Assay Information

Assay Type:

Biochemical

Assay Sub Type:

Binding

Detection Method:

Radiometry

Measured Response:

Scintillation

Testing Information

Procedure Summary:

This assay measures the binding of [³H]BNZA to voltage-gated sodium channel Nav1.5. Human recombinant Nav1.5 expressed in CHO cells is used in modified Tris-HCl buffer pH 7.4. A 20 µg‡ aliquot of membrane is incubated with 1 nM [³H]BNZA for 180 minutes at 25℃. Non-specific binding is estimated in the presence of 100 µM Dibucaine. Membranes are filtered and washed 4 times, and the filters are counted to determine [³H]BNZA specifically bound. Compounds are screened at 10 µM.
Note: ‡Membrane protein may change from lot to lot, the concentration used will be adjusted if necessary.

Ligand:

[³H]BNZA

Ligand Kd (nM):

11

Ligand Concentration:

1 nM

Non Specific:

100 µM Dibucaine

Incubation:

180 min at 25°C

Control Inhibitor:

Dibucaine

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:

2

Turnaround Time

Standard:

10 Business Days

Reference Compound Data

PubChem ID

6610253

Name

Veratridine

Measurement

>10μM - IC50

PubChem ID

132988485

Name

Saxitoxin

Measurement

>1μM - IC50

PubChem ID

441074

Name

Quinidine

Measurement

3.5μM - IC50

PubChem ID

521951

Name

Dibucaine

Measurement

0.49μM - IC50

PubChem ID

6419868

Name

Aconitine

Measurement

8.7μM - IC50

Clinical Relevance

Therapeutic Area:

Cardiovascular

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei

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