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Screen Quest™ No Wash Potassium Channel Assay Kit
Potassium (K+) ion channel plays an important role in regulating fundamental biological processes including heart rate, hormone and neurotransmitter secretion, water and electrolyte balance. Potassium ion channel has been considered as drug targets for disease indications including arrhythmia, pain, diabetes, neurological dysfunctions etc. The permeability of Tl+ through K+ channel has been widely used to assay K+ channel. The cells that express K+ channel of interests (e.g. hERG, Kv1.3, Kir2.1, KATP) are pre-loaded with a Tl+ sensitive dye. The dye is non-fluorescent and is permeable to cell membrane. Once inside the cell, the non-fluorescent AM ester dye is cleaved by endogenous esterase into a negatively charges dye that stays inside cells. When a stimulus buffer containing low dose of Tl+ is added to cells, the Tl+ flows across the K+ channel and binds to Tl+ sensitive dye, generating a fluorescent signal. This signal is proportional to the activity of K+ channel. If an antagonist or antagonist is added to the cells, the fluorescent signal decreases or increases respectively, to reflect the inhibited or stimulated activity of K+ channel.
Astemizole dose dependent inhibition of hERG channel was measured in HEK293-KCNH2 cells with Screen Quest™ Potassium Ion Channel Kit. The cells were seeded overnight at 20,000 cells/100 µL/well in a Costar black wall/clear bottom 96-well poly-D-lysine plate. The cells were incubated with 100 µL of dye-loading solution for 1 hour at 37°C. 10 µL of Astemizole was added to the cells and incubated for 30 minutes at 37°C. 50 µL of 0.5 mM Tl2SO4 and 2.5 mM K2SO4 containing stimulus solution was injected in each well by FlexStation and read every sec for 3 minutes at excitation/emission=490/525nm. IC50 = 0.46 µM.
Astemizole dose dependent inhibition of hERG channel was measured in HEK293-KCNH2 cells with Screen Quest™ Potassium Ion Channel Kit. The cells were seeded overnight at 20,000 cells/100 µL/well in a Costar black wall/clear bottom 96-well poly-D-lysine plate. The cells were incubated with 100 µL of dye-loading solution for 1 hour at 37°C. 10 µL of Astemizole was added to the cells and incubated for 30 minutes at 37°C. 50 µL of 0.5 mM Tl2SO4 and 2.5 mM K2SO4 containing stimulus solution was injected in each well by FlexStation and read every sec for 3 minutes at excitation/emission=490/525nm. IC50 = 0.46 µM.
protocol
Protocol
COA
CatalogSize
Price
Quantity
365501 Plate
Price
3655110 Plates
Price
36552100 Plates
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Spectral properties
Excitation (nm)494
Emission (nm)516
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200
Instrument settings
Fluorescence microplate reader
Excitation490 nm
Emission525 nm
Cutoff515 nm
Recommended plateBlack wall/clear bottom
Instrument specification(s)Bottom read mode/Programmable liquid handling
Other instrumentsArrayScan, FDSS, FLIPR, FlexStation, IN Cell Analyzer, NOVOStar, ViewLux
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Page updated on October 13, 2025