What do you mean with relative fluorescent units (immunoblot, FACS, plate readout)?
Ok so for this you need several things.
A good fluorochrome (e.g. H2DCFDA, DHE, MitoSox, etc. depending on your experimental goals). Additionally, you need the right reader which can detect fluorescence with the correct filters regarding the dye you want to use.
But the most crucial step is the experimental conditions.
So what do you want to look for? Stimulation of some ligands? Effects of drugs etc.?
Dear Sir,
I am working in MIN6 cells, i want to do experiment to measure Calcium influx for that i am using FURA2 as a fluorescent dye. for this measurement i want to calculate the fluorescent intensity in RFU units .
Please give me suggestion
Ok these are the informations I wanted to know. As i read MIN6 are adherent cells, right?
For calcium flux measurements you should keep in mind some very important informations. fura2 should be skipped to its esterized form fura2-am. this form can be internilized and desterized intracellulary. so you should also have a look for the concentration and incubation time. so it also depends on the reading method (plate reader, FACS, microscopy) and the stimulation/modulation/inhibition etc. just see the two publications below.
http://www.nature.com/ncb/journal/v1/n3/pdf/ncb0799_165.pdf
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