Development and validation of HTS flux assay for endogenously expressed chloride channels in a CHO-K1 cell line

Assay and Drug Development Technologies
Sikander GillDong Liang

Abstract

An atomic absorption spectroscopy-based detection system was employed to develop a new non-radioactive flux assay for chloride (Cl-) channels in a high throughput format. Cl- flux is assayed by measuring the extent to which Cl- precipitates an excess amount of silver ions (Ag+). A linear correlation was observed between theoretical and determined Cl- concentration with an r2 value of 0.996. The assay was found to be free from interference from various ions and proteins. The assay was used to study the physiology of endogenously expressed Cl- channels in a Chinese hamster ovary-K1 cell line. Cl- efflux was activated in response to an increased concentration of K+ (100 mM), Ca2+ (4 mM), and ionomycin (10 microM) as calcium ionophore. The efflux was also sensitive to pH as slightly higher efflux of Cl- was observed at an acidic pH of 3.2 in comparison to the neutral pH of 7.4. The Cl- efflux was inhibited by 100 microM 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) and 500 microM 5-nitro-2-(3-phenylpropylamino) benzoate (NPPB) but not by tolbutamide, niflumic acid, or glybenclamide, indicating that the channel current is not sensitive to other cystic fibrosis transmembrane conductance regulator inhibitors. The 50% inhibit...Continue Reading

References

Feb 1, 1992·Pflügers Archiv : European journal of physiology·G Wu, O P Hamill
Sep 1, 1992·The American Journal of Physiology·S E GabrielM J Stutts
Nov 15, 1993·The Biochemical Journal·M J Mason, S Grinstein
Jun 6, 2000·Journal of Biomolecular Screening·J H ZhangK R Oldenburg
Jun 9, 2000·Annual Review of Physiology·J F Kidd, P Thorn
Aug 15, 2000·Annual Review of Biophysics and Biomolecular Structure·M MadukeJ A Mindell
May 25, 2001·Analytical Biochemistry·C D GeddesD J Birch
Oct 16, 2001·American Journal of Physiology. Cell Physiology·L V GaliettaA S Verkman
Mar 28, 2002·Physiological Reviews·Thomas J JentschAnselm A Zdebik
Oct 12, 2004·Journal of Biomolecular Screening·Weimin Tang, Mary Jo Wildey
Jan 27, 2005·Assay and Drug Development Technologies·Kewei WangJohn Dunlop
Jan 27, 2005·Assay and Drug Development Technologies·Wei ZhengLaszlo Kiss
Jun 25, 2005·Current Pharmaceutical Design·J A ArmijoJ Adín

❮ Previous
Next ❯

Citations

Dec 14, 2011·Nihon yakurigaku zasshi. Folia pharmacologica Japonica·Masato FujiiYuji Imaizumi
Mar 24, 2011·Future Medicinal Chemistry·Georg C TerstappenRavikumar Peri
Nov 10, 2012·Journal of Pharmacological and Toxicological Methods·Katsutoshi IdoKohei Sawada
May 9, 2013·Journal of Biomolecular Screening·Rangan MaitraKeith Warner
Jun 21, 2017·Assay and Drug Development Technologies·Sikander GillDong Liang

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.