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Applications: Drug compound screening for activators or inhibitors of IKCA1
Functional assays for IKCA1
Efficient antigen for mouse immunization
Background: IKCA1 is part of a potentially heterotetrameric voltage-independent potassium channel that is activated by intracellular calcium. Activation is followed by membrane hyperpolarization, which promotes calcium influx. The encoded protein may be part of the predominant calcium-activated potassium channel in T-lymphocytes.
Biological Activity: N-terminal FLAG-tagged human IKCA1 has been stably expressed in HEK293 cell line and its expression was confirmed by Western blotting. Functional analysis of IKCA1 channel in the stable recombinant HEK293 was accomplished by using the membrane potential-sensitive fluorescence dye DiBAC4(3). When IKCA1-expressed HEK293 cells were stimulated with calcium-specific ionophore ionomycin to raise the intracellular level of Ca2+, IKCA1 channel was activated, leading to hyperpolarization and a decrease in cell fluorescence in the functional fluorescence assay with DiBAC4(3). This decrease of cell fluorescence was blocked by the IKCA1 channel inhibitor, clotrimazole (CLT).
Description: Stable, recombinant HEK293 cell line expressing human IKCA1, also known as KCNN4 (Intermediate conductance calcium-activated potassium channel protein 4), IK1, hKCa4, and hSK4, GenBank Accession No. NM_002250.
Genbank: NM_002250
Host Cell Line: HEK293
Mycoplasma Testing: The cell line has been screened using the PCR-based Venor™GeM Mycoplasma Detection kit (Sigma Aldrich) to confirm the absence of Mycoplasma species.
Storage Stability: Store in liquid nitrogen immediately upon receipt.
Supplied As: Each vial contains ~2 X 10^6 cells in 1 ml of 10% DMSO.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: BSL-2
References: 1. Ghanshani S et al. (2000). J Biol Chem. 275(47): 37137-37149.
2. Hoffman JF et al. (2003). PNAS 100 (12): 7366-7371.
3. de-Allie FA et al. (1996). Br J Pharmacol. 117(3):479-487.
4. Terstappen GC et al. (2003). Neurosci Lett. 346(1-2):85-88.
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