Improvements in Live Cell Analysis of G Protein Coupled Receptors using Second Generation BD Calcium Assay Kits
نویسندگان
چکیده
BD Calcium Assay Kits are designed for cell-based calcium mobilization high-throughput screening assays. The kits use a proprietary formulation including a non-fluorescent calcium indicator that becomes activated inside the cell and shows increased fluorescence upon calcium binding. The formulation includes a signal-enhancing reagent to maximize the signal over background in a homogeneous, no-wash assay format, based on a technology developed at BD. We have compared the next generation BD calcium assay kit product family to previous versions of the formulation, and to other commercially available homogeneous calcium assay kits. The improvements have enabled better performance on the cell lines and receptors that we have tested in all plate formats including 1536.
منابع مشابه
High glucose condition down-regulates the inhibitory G-protein subunit, Gαi, in pheochromocytoma PC12 cells
Introduction: G-proteins have an important role in the cell signaling of numerous receptors. The situation of G-proteins in health and disease and their critical role in the development of diabetic side effects is an interested scientific field. Here, the changes in the expression of G-protein subunits (Gαi, Gαs and Gβ) were evaluated in hyperglycemic situation of PC12 cells as...
متن کاملPossible Involvement of a Specific Cell Surface Receptor for Calprotectin-Induced Apoptosis in Colon Adenocarcinoma and Carcinam Cell Lines (SW742 and HT29/219)
Calprotectin, a calcium-bound protein complex, is abundant in the cytosol of neutrophils. It has been reported that this protein has an apoptotic activity in tumor cells. Since calprotectin increases in colorectal cancer, this study was conducted to investigate, for the first time, the cytotoxicity/apoptotic effect of calprotectin on HT29/219 and SW742 colon carcinoma and adenocarcinoma cell li...
متن کاملEvaluation of FLIPR Calcium 3 Assay Kit--a new no-wash fluorescence calcium indicator reagent.
We have evaluated the FLIPR Calcium 3 Assay Kit (Calcium 3), a new no-wash fluorescence calcium indicator dye reagent, for the measurement of agonist-stimulated calcium signaling in cells expressing the serotonin 2C (5-HT(2C)), metabotropic glutamate receptor 5 (mGluR5) and the vasopressin 2 (V2) G-protein-coupled receptors. Calcium 3 yielded equivalent (5-HT(2C)) or superior (mGluR5 and V2) se...
متن کاملChimeric G proteins extend the range of insect cell-based functional assays for human G protein-coupled receptors.
We previously described a functional assay for G protein-coupled receptors (GPCRs) based on stably transformed insect cells and using the promiscuous G protein Galpha16. We now show that, compared with Galpha16, the use of chimeric Galphaq subunits with C-terminal modifications (qi5-HA, qo5-HA, or qz5-HA) significantly enhances the ability of insect cells to redirect Gi-coupled GPCRs into a Gq-...
متن کاملFunctional analysis of endogenous beta-adrenergic receptor through fluorimetric monitoring of cyclic nucleotide-gated ion channel.
b-Adrenergic receptors (b1, b2, and b3), coupling either to heterotrimeric G protein Gs solely or to Gs and Gi [1,2], regulate the activity of adenylyl cyclase that controls the intracellular level of cyclic 3 0,5 0-adenosine monophosphate (cAMP). Currently, many cAMP assays are used to analyze endogenous b-adrenergic receptor function. However, most of the existing cAMP assays for G protein-co...
متن کامل