Ionomycin is a n antibiotic that acts as a potent and selective calcium ionophore. Cell signaling technology 3 trask lane danvers, ma 0 923 usa 9788672300 tel 9788672400 fax 9785786737 emergency tel ii. Fura2 free acid is then added to each solution to achieve a final concentration of 100. The data acquisition program used metafluor, imagej, or an. Ionomycin calcium salt supplier cas 56092821 tocris. Selected citations for ionomycin calcium salt include. Each solution also includes 50 m fura2, as well as 15 m. Ionomycin calcium salt ready made solution, from streptomyces. Kindly submitted by prof amy harkins, st louis university. The fura2 calcium imaging calibration kit is designed to facilitate rapid calibration and standardization of digital imaging microscopes. It is also known for its antiproliferative effects. Calibration of calcium levels 1 tools for cardiovascular research contractility and ratiometric fluorescence systems calibration of calcium levels the ratiometric method of determining cell calcium levels using fura2 is simple and elegant.
You can use it to display info about returns and refunds, latest promotions etc. It is produced by the bacterium streptomyces conglobatus. Ionomycin calcium salt from streptomyces conglobatus catalog number i0634 storage temperature 28 c cas rn. Ionomycin induces apoptosis in immature b cell lines and in cultured embryonic rat cortical neurons. Calibrating indicators within organelles is often more challenging than. In cultured chicken pectoralis muscle, ionomycin 106105 m selectively increased the incorporation of methionine into two proteins. Measurement of intracellular calcium concentration. Material safety data sheet msds for ionomycin, calcium salt i. Loading and calibration of intracellular ion indicatorsnote 19. Ionomycin, calcium salt ionomycin is commonly used to modify intracellular calcium concentrations and to calibrate fluorescent calcium indicators. These measurements and the calibration of fura2 fluorescence was performed according to standard protocols. Ionomycin calcium salt ready made solution, from streptomyces conglobatus, 1 mm in dmso, 0. In vivo calibration requires a combination of patch clamping and calcium imaging, which can be complicated but is also very precise. How does a calcium imaging calibration for fura2 look like.
Calcium imaging is a valuable tool to study the regulation of intracellular. I have been looking at the flipr calcium assay kits from molecular devices. Ionomycin calcium salt 56092821 buy ionomycin calcium. The fluorescence of the indicator must be quantitatively related to the.
Ionomycin is a potent and selective calcium ionophore derived from streptomyces conglobatus liu et al. Citations are publications that use tocris products. Ionomycin is a selective calcium ionophore derived from s. This protocol has been successfully adapted for different cell types, including fibroblasts, pc12 cells and embryonic neurons with variations on cell density and fura2 am concentration. Measuring calcium dynamics in living cells with genetically. It was found that the antibiotic is capable of extracting calcium ion from the bulk of an aqueous phase into an organic phase. Post calibration at the end of each experiment can be done using ionomycin. Take the hbss and 50 mm k solutions out of the refrigerator, turn on equipment, perform calibration curve, check. In vivo calibration requires a combination of patch clamping and calcium. Actions of ionomycin, 4bra23187 and a novel electrogenic. Ionomycin is a potent and selective calcium ionophore agent 1,2. Ionomycin is a very longchain fatty acid that is docosa10,16dienoic acid which is substituted by methyl groups at positions 4, 6, 8, 12, 14, 18 and 20, by hydroxy groups at positions 11, 19 and 21, and by a 2,5dimethyloctahydro2,2bifuran5ylethanol group at position 21. Apr 29, 2019 calcium imaging calibration posted in pharmacology and pharmaceutics.
Intracellular calibration of the fluorescent calcium. A method for estimating intracellular ion concentration. Calcium imaging calibration pharmacology and pharmaceutics. A j morgan and r jacob vascular biology research centre, kings college london, university of london, kensington, u. I want to perform a calibration for calcium imaging experiments and. In vitro calibration can be done using a homemade calibration chamber that consists of two coverslips separated by a thin coverslip spacer to generate a thin layer of solution in which to perform the calibration. One measures the fluorescence emission excited by two separate wavelengths. One measures the fluorescence emission excited by two separate wavelengths, subtracts the background signal and the ratio between the intensity levels is used to calculate the free calcium concentration. Fura2 calcium imaging calibration kit life technologies, invitrogen, catalog.
We enable science by offering product choice, services, process excellence and our people make it happen. See actual entry in rtecs for complete information. The ionophorous properties of a new antibiotic, ionomycin, have been studied. Fura2 am calcium imaging protocol for fibroblasts, pc12, embryonic neurons and more.
Chelators, calibration buffers, ionophores and cell. How can i use ionomycin and egta to do in situ calibration of fura2am for calcium imaging. Ionomycin is an ionophore produced by the bacterium streptomyces conglobatus. Calibration of calcium levels the ratiometric method of determining cell calcium levels using fura2 is simple and elegant.
Only select registry of toxic effects of chemical substances rtecs data is presented here. If you want absolute levels of calcium, you have to calibrate your experimental setup by chelating all calcium with egta to substract background fluorescence and known amounts of calcium make. The kd of an indicator can be calculated from a plot generated by scanning the excitation or emission of the indicator. To examine whether ionomycin could stimulate influx via a storedependent mechanism. Zeiss inverted microscopy with perfusion system and iplab software. Chelators, calibration buffers, ionophores and cellloading. Free calcium concentrations in lclc 103h cells were determined before and after adding 2. Which would be the proper concentrations of ca and egta in order to get a transient response curve within 60 seconds. Actions of ionomycin, 4bra23187 and a novel electrogenic ca2. The antibiotic also acts as a mobile ion carrier to transport the cation across a solvent barrier. Calcium ionomycin product description molecular formula.
Measurement of intracellular calcium concentration in. Alternatively, premade solutions are available as a calcium calibration buffer kit from invitrogen. Ionomycin calcium salt investigated as a drug and natural product. The effect of ionomycin and a23187 on the mitochondrial potential in intact cells is less clear. Characterization of ionomycin as a calcium ionophore. Calcium calibration buffer kits thermo fisher scientific. Ionomycin calcium salt calcium ionophore hello bio.
Fay2 i department of physiology, university of melbourne, parkvife, victoria, australia 2 department of physiology, university of massachusetts medical school, worcester, massachusetts, usa abstract we present the. Usa software that converts the ratiometric signal into an actual calcium read out if. Ionomycin, calcium salt, streptomyces conglobatus, cas 56092821, is useful in cell activation experiments when calcium doseresponse data are not required. Calcium imaging calibration posted in pharmacology and pharmaceutics. Calcium imaging of cortical neurons using fura2 am protocol. Antibiotic that acts as a potent and selective calcium ionophore.
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