PMC:4440627 / 11393-12215 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/4440627","sourcedb":"PMC","sourceid":"4440627","source_url":"https://www.ncbi.nlm.nih.gov/pmc/4440627","text":"Ca2+ Flux Measurements\nCytosolic Ca2+ Flux was measured using flow cytometry. Breifly, cells were loaded for 30 min at 37°C with 2μM indo-1 acetoxylmethylester (Indo-1 AM) (Molecular Probes) in Hanks’ balanced salt solution (HBSS) (Invitrogen Cat # 14025) with 1% FCS and 2.5mM probecinide (Invitrogen). Cells were washed once and resuspended in calcium and magnesium free HBSS (Invitrogen # 14175) with 1% FCS. To measure intracellular store calcium flux cells were treated with 1mM EGTA and subsequently with thapsigargin, ionomycin, or m-3m3FBS (Tocris Bioscience) and changes in fluorescence intensity were monitored on an LSRII flow cytometer (BD Biosciences). Values were plotted as the ratio of fluorescence at Ca2+-bound Indo-1 AM to that of Ca2+-free Indo-1 AM. Data were analyzed with FlowJo software (TreeStar).","divisions":[{"label":"Title","span":{"begin":0,"end":22}}],"tracks":[]}