Store-operated calcium cellular assay
The present technology provides a cell based assay for identifying compounds that modulate store-operated ionic calcium levels using itpr mutant cell lines, such as itpr-ku cells, which have abnormal ionic calcium levels.
1. A kit for identifying a compound that modulates store-operated calcium entry (SOCE) level in a cell, comprising:
at least one mutant mammalian and/or avian cell, wherein the mutant mammalian and/or avian cell comprises a residual SOCE level and expresses a single 1,4,5-inositol triphosphate receptor (InsP 3 R) gene (itpr) product wherein the itpr gene encoding the single InsP 3 R gene product consists of an itpr ka1091/ug3 mutant, and wherein the itpr ka1091/ug3 mutant contains G1891S and S224F point mutations; and at least one control cell.
2. The kit of claim 1 , wherein the control cell is selected from the group consisting of: (i) a wild-type, normal cell, and (ii) a cell having wild-type calcium ion release.
3. The kit of claim 1 , wherein the mutant mammalian cell or the mutant avian cell is selected from the group consisting of a 3T3 cell, a BHK21 cell, an MDCK cell, a HeLa cell, a PtK1 cell, an L6 cell, a PC12 cell, an SP2 cell, a COS cell a 293 cell, a CHO cell, a DT40 cell, an R1 cell, and E14.1 cell, an H1 cell, and an H9 cell.
4. The kit of claim 1 , further comprising a component for calcium imaging.
5. The kit of claim 4 , wherein the component for calcium imaging comprises a fluorescent dye.
6. The kit of claim 5 , wherein the fluorescent dye comprises fluo-4.
7. The kit of claim 1 , further comprising thapsigargin.
8. The kit of claim 1 , wherein the mutant mammalian cell is a murine cell.