new BindingDB logo
myBDB logout

Assay Method Information

Assay Name:  Electrophysiological Assay
Description:  Current record was obtained by an automated patch clamp system “IonWorks Quattro (Molecular Devices Corporation)” in Population Patch Clamp mode. The operation was conducted in accordance with the operating procedure of the system. A Dulbecco's phosphate buffer containing calcium and magnesium (Sigma) was used as an extracellular fluid, and a low C1-buffer (100 mM K-gluconate, 40 mM KCl, 3.2 mM MgCl2, 5 mM EGTA, 5 mM Hepes, pH 7.3) was used as an intracellular fluid. A test compound was dissolved in dimethylsulfoxide (DMSO) to prepare a 30 mM stock solution, so as to produce 4-fold serial dilutions with the extracellular fluid for attaining a DMSO concentration of 0.3% in measurement.The hNav 1.7/β1/β2 cells cultured to a 70-80% confluent state in a T150 flask (Sumilon) were washed with PBS and subsequently with versene (Invitrogen Corp.), and collected by allowing to react with 0.05% trypsin (Invitrogen Corp.) at 37° C. for 3 minutes. After washing with a culture medium, the resultant cells were suspended in an extracellular fluid at a concentration of 2×10−6 cells/ml so as to be used for the measurement. The cell membrane was perforated by using an intracellular fluid including 100 μg/ml amphotericin B (Sigma).
Affinity data for this assay
 

If you find an error in this entry please send us an E-mail
   
    

Home

|

Search

|

Deposit

|

SiteMap

|

About us

|

Email us

|

Info

 
Last update November 1, 2007
©2000 BindingDB. All rights reserved.