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CAS:112926-02-0
Appearance:Solid
Storage:Store powder at -20℃ for 2 years, in solvent(mother liquid) at -20℃ for 1 month, -80℃ for 6 months.
Purity:≥95%
Manual Download
Calcium measurements are critical for numerous biological investigations. Fluorescent probes that show spectral responses upon binding to Ca2+ have enabled researchers to investigate changes in intracellular free Ca2+ concentrations by using fluorescence microscopy, flow cytometry, fluorescence spectroscopy and fluorescence microplate readers. This cell-permeant Indo-1 AM, is a UV light excitable, emission ratioable Ca2+ indicator. Upon binding to Ca2+, the emission maximum of Indo-1 AM shifts from 480 nm to 400 nm. Indo-1 is preferred for flow cytometry, in which it is more practical to use a single laser for excitation, such as the 351-364 nm spectral lines of the argon-ion laser.
Fluorescence microscope（for reference only）
Excitation Indo-1 filter set
Emission Indo-1 filter set
Recommended plate Black wall/clear bottom
Fluorescence microplate reader（for reference only）
Excitation 340
Emission 400, 475
Cutoff Ex/Em = 340/400, no cut off. Ex/Em = 340/475, cut off 455
Recommended plate Black wall/clear bottom
Instrument specification(s) Bottom read mode/Programmable liquid handling
Protocol (for reference only)
Preparative storage solution
Dilute Indo-1 AM with DMSO to prepare a 10 mM reserve solution.
Note:
Indo-1 AM storage solution is recommended to be stored away from light at -20 ℃ or -80 ℃ after packaging.
Preparation of working fluid
The storage solution was diluted with preheated HBSS to prepare 1-10 μM Indo-1 AM working solution.
Note:
Please adjust the concentration of Indo-1 AM working fluid according to the actual situation. Use immediately after dissolution.
Cell staining
Suspension cell
1 Centrifuge the cells and wash them twice with PBS for 5 minutes each time. The cell density was 1×106/mL.
2 Add 1 mL of Indo-1 AM working liquid and incubate at room temperature for 5-30 minutes.
3 400 xg, 3-4 minutes, discard the supernatant.
4 Add PBS to wash cells twice for 5 minutes each time.
5 Cells were suspended with 1 mL HBSS and observed using fluorescence microscopy or flow cytometry.
Adherent cell
1 The adherent cells were cultured on a sterile cover slide.
2 Remove the cover glass from the medium and absorb the excess medium.
3 Add 100 μL dye working solution, shake gently to completely cover the cells, incubate for 5-30 minutes.
4. Remove the dye working solution, wash it with the medium 2-3 times for 5 minutes each time, and observe it with a fluorescence microscope.
