Indo-1, AM *CAS 112926-02-0*
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
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International | See distributors |
Shipping | Standard overnight for United States, inquire for international |
Physical properties
Dissociation constant (Kd, nM) | 230 |
Molecular weight | 1009.91 |
Solvent | DMSO |
Spectral properties
Extinction coefficient (cm -1 M -1) | 33000 |
Excitation (nm) | 330 |
Emission (nm) | 404 |
Storage, safety and handling
Certificate of Origin | Download PDF |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
Storage | Freeze (< -15 °C); Minimize light exposure |
UNSPSC | 12352200 |
Alternative formats
Indo-1, AM *UltraPure Grade* *CAS 112926-02-0* |
Overview | ![]() ![]() |
See also: Calcium Indicators, Intracellular Ions, Physiological Probes, Ratiometric Calcium Indicators
CAS 112926-02-0 | Molecular weight 1009.91 | Dissociation constant (Kd, nM) 230 | Extinction coefficient (cm -1 M -1) 33000 | Excitation (nm) 330 | Emission (nm) 404 |
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.
Platform
Fluorescence microscope
Excitation | Indo-1 filter set |
Emission | Indo-1 filter set |
Recommended plate | Black wall/clear bottom |
Fluorescence microplate reader
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 |
Example protocol
PREPARATION OF STOCK SOLUTIONS
Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
Indo-1 AM Stock Solution
Prepare a 2 to 5 mM stock solution of Indo-1 AM in high-quality, anhydrous DMSO. PREPARATION OF WORKING SOLUTION
Indo-1 AM Working Solution
On the day of the experiment, either dissolve Indo-1 AM in DMSO or thaw an aliquot of the indicator stock solution to room temperature. Prepare a dye working solution of 2 to 20 µM in a buffer of your choice (e.g., Hanks and Hepes buffer) with 0.04% Pluronic® F-127. For most cell lines, Indo-1 AM at a final concentration of 4-5 μM is recommended. The exact concentration of indicators required for cell loading must be determined empirically.Note The nonionic detergent Pluronic® F-127 is sometimes used to increase the aqueous solubility of Indo-1 AM. A variety of Pluronic® F-127 solutions can be purchased from AAT Bioquest.
Note If your cells contain organic anion-transporters, probenecid (1-2 mM) may be added to the dye working solution (final in well concentration will be 0.5-1 mM) to reduce leakage of the de-esterified indicators. A variety of ReadiUse™ probenecid products, including water-soluble, sodium salt, and stabilized solution, can be purchased from AAT Bioquest.
SAMPLE EXPERIMENTAL PROTOCOL
Following is our recommended protocol for loading AM esters into live cells. This protocol only provides a guideline and should be modified according to your specific needs.
- Prepare cells in growth medium overnight.
- On the next day, add 1X Indo-1 AM working solution into your cell plate.
Note If your compound(s) interfere with the serum, replace the growth medium with fresh HHBS buffer before dye-loading. - Incubate the dye-loaded plate in a cell incubator at 37 °C for 30 to 60 minutes.
Note Incubating the dye for longer than 1 hour can improve signal intensities in certain cell lines. - Replace the dye working solution with HHBS or buffer of your choice (containing an anion transporter inhibitor, such as 1 mM probenecid, if applicable) to remove any excess probes.
- Add the stimulant as desired and simultaneously measure fluorescence using either a fluorescence microscope equipped with an Indo-1 filter set or a fluorescence plate reader containing a programmable liquid handling system such as a FlexStation, at Ex/Em1 = 340/400 nm no cutoff and Ex/Em2 = 340/475 cutoff 455 nm.
Calculators
Common stock solution preparation
Table 1. Volume of DMSO needed to reconstitute specific mass of Indo-1, AM *CAS 112926-02-0* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 99.019 µL | 495.094 µL | 990.187 µL | 4.951 mL | 9.902 mL |
5 mM | 19.804 µL | 99.019 µL | 198.037 µL | 990.187 µL | 1.98 mL |
10 mM | 9.902 µL | 49.509 µL | 99.019 µL | 495.094 µL | 990.187 µL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
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Spectrum
Open in Advanced Spectrum Viewer


Spectral properties
Extinction coefficient (cm -1 M -1) | 33000 |
Excitation (nm) | 330 |
Emission (nm) | 404 |
Images
Citations
View all 5 citations: Citation Explorer
Development of a deep two-photon calcium imaging method for the analysis of cortical processing in the mammalian brain
Authors: Birkner, Antje
Journal: (2019)
Authors: Birkner, Antje
Journal: (2019)
Nifedipine stimulates proliferation and migration of different breast cancer cells by distinct pathways
Authors: Zhao, Tao and Guo, Dongqing and Gu, Yuchun and Ling, Yang
Journal: Molecular Medicine Reports (2017): 2259--2263
Authors: Zhao, Tao and Guo, Dongqing and Gu, Yuchun and Ling, Yang
Journal: Molecular Medicine Reports (2017): 2259--2263
Inhibition of pyruvate kinase M2 by reactive oxygen species contributes to the development of pulmonary arterial hypertension
Authors: Guo, Dongqing and Gu, Junzhong and Jiang, Hui and Ahmed, Asif and Zhang, Zhiren and Gu, Yuchun
Journal: Journal of molecular and cellular cardiology (2016): 179--187
Authors: Guo, Dongqing and Gu, Junzhong and Jiang, Hui and Ahmed, Asif and Zhang, Zhiren and Gu, Yuchun
Journal: Journal of molecular and cellular cardiology (2016): 179--187
Nifedipine promotes the proliferation and migration of breast cancer cells
Authors: Guo, Dong-Qing and Zhang, Hao and Tan, Sheng-Jiang and Gu, Yu-Chun
Journal: PloS one (2014): e113649
Authors: Guo, Dong-Qing and Zhang, Hao and Tan, Sheng-Jiang and Gu, Yu-Chun
Journal: PloS one (2014): e113649
References
View all 84 references: Citation Explorer
Measurement of [Ca2+] in cell suspensions using indo-1
Authors: Nelemans A., undefined
Journal: Methods Mol Biol (2006): 47
Authors: Nelemans A., undefined
Journal: Methods Mol Biol (2006): 47
A flow cytometric comparison of Indo-1 to fluo-3 and Fura Red excited with low power lasers for detecting Ca(2+) flux
Authors: Bailey S, Macardle PJ.
Journal: J Immunol Methods (2006): 220
Authors: Bailey S, Macardle PJ.
Journal: J Immunol Methods (2006): 220
Ratiometric intracellular calcium imaging in the isolated beating rat heart using indo-1 fluorescence
Authors: Eerbeek O, Mik EG, Zuurbier CJ, van 't Loo M, Donkersloot C, Ince C.
Journal: J Appl Physiol (2004): 2042
Authors: Eerbeek O, Mik EG, Zuurbier CJ, van 't Loo M, Donkersloot C, Ince C.
Journal: J Appl Physiol (2004): 2042
Negative inotropic effects of angiotensin II, endothelin-1 and phenylephrine in indo-1 loaded adult mouse ventricular myocytes
Authors: Sakurai K, Norota I, Tanaka H, Kubota I, Tomoike H, Endo M.
Journal: Life Sci (2002): 1173
Authors: Sakurai K, Norota I, Tanaka H, Kubota I, Tomoike H, Endo M.
Journal: Life Sci (2002): 1173
Usefulness of the analytic method of intracellular calcium and the problems--aequorin and indo-1 signal
Authors: Endoh M., undefined
Journal: Nippon Yakurigaku Zasshi (2000): 361
Authors: Endoh M., undefined
Journal: Nippon Yakurigaku Zasshi (2000): 361
Comment on "Usefulness of intracellular calcium analysis and the problem--aequorin and indo-1 signal"
Authors: Imaizumi Y., undefined
Journal: Nippon Yakurigaku Zasshi (2000): 101
Authors: Imaizumi Y., undefined
Journal: Nippon Yakurigaku Zasshi (2000): 101
Concentrations of caffeine greater than 20 mM increase the indo-1 fluorescence ratio in a Ca(2+)-independent manner
Authors: McKemy DD, Welch W, Airey JA, Sutko JL.
Journal: Cell Calcium (2000): 117
Authors: McKemy DD, Welch W, Airey JA, Sutko JL.
Journal: Cell Calcium (2000): 117
Intracellular calcium signals measured with indo-1 in isolated skeletal muscle fibres from control and mdx mice
Authors: Collet C, Allard B, Tourneur Y, Jacquemond V.
Journal: J Physiol (1999): 417
Authors: Collet C, Allard B, Tourneur Y, Jacquemond V.
Journal: J Physiol (1999): 417
Measurement of [Ca2+]i in cell suspensions using indo-1
Authors: Nelemans A., undefined
Journal: Methods Mol Biol (1999): 41
Authors: Nelemans A., undefined
Journal: Methods Mol Biol (1999): 41
Alpha-stat calibration of indo-1 fluorescence and measurement of intracellular free calcium in rat ventricular cells at different temperatures
Authors: Wang SQ, Zhou ZQ.
Journal: Life Sci (1999): 871
Authors: Wang SQ, Zhou ZQ.
Journal: Life Sci (1999): 871
Application notes
What's A Ratiometric Indicator
A Comparison of Fluorescent Red Calcium Indicators for Detecting Intracellular Calcium Mobilization in CHO Cells
A Meta-Analysis of Common Calcium Indicators
A New Red Fluorescent & Robust Screen Quest™ Rhod-4™ Ca2+Indicator for Screening GPCR & Ca2+ Channel Targets
A New Robust No-Wash FLIPR Calcium Assay Kit for Screening GPCR and Calcium Channel Targets
A Comparison of Fluorescent Red Calcium Indicators for Detecting Intracellular Calcium Mobilization in CHO Cells
A Meta-Analysis of Common Calcium Indicators
A New Red Fluorescent & Robust Screen Quest™ Rhod-4™ Ca2+Indicator for Screening GPCR & Ca2+ Channel Targets
A New Robust No-Wash FLIPR Calcium Assay Kit for Screening GPCR and Calcium Channel Targets