Fluo-8H™, AM
Price | |
Catalog Number | |
Unit Size | |
Quantity |
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Dissociation constant (Kd, nM) | 232 |
Molecular weight | 1074.98 |
Solvent | DMSO |
Correction Factor (260 nm) | 1.076 |
Correction Factor (280 nm) | 0.769 |
Extinction coefficient (cm -1 M -1) | 23430 |
Excitation (nm) | 495 |
Emission (nm) | 516 |
Quantum yield | 0.161 |
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 |
Overview | ![]() ![]() |
Molecular weight 1074.98 | Dissociation constant (Kd, nM) 232 | Correction Factor (260 nm) 1.076 | Correction Factor (280 nm) 0.769 | Extinction coefficient (cm -1 M -1) 23430 | Excitation (nm) 495 | Emission (nm) 516 | Quantum yield 0.161 |
Platform
Fluorescence microscope
Excitation | FITC |
Emission | FITC |
Recommended plate | Black wall/clear bottom |
Fluorescence microplate reader
Excitation | 490 |
Emission | 525 |
Cutoff | 515 |
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
Prepare a 2 to 5 mM stock solution of Fluo-8H™ AM in high-quality, anhydrous DMSO.
PREPARATION OF WORKING SOLUTION
On the day of the experiment, either dissolve Fluo-8H™ AM in DMSO or thaw an aliquot of the indicator stock solution to room temperature.
Prepare a 2 to 20 µM Fluo-8H™ AM working solution in a buffer of your choice (e.g., Hanks and Hepes buffer) with 0.04% Pluronic® F-127. For most cell lines, Fluo-8H™ 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 Fluo-8H™ 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 solutions, 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 Fluo-8H™ AM working solution to 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 2 hours 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 a FITC filter set or a fluorescence plate reader containing a programmable liquid handling system such as an FDSS, FLIPR, or FlexStation, at 490/525 nm cutoff 515 nm.
Calculators
Common stock solution preparation
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 93.025 µL | 465.125 µL | 930.25 µL | 4.651 mL | 9.302 mL |
5 mM | 18.605 µL | 93.025 µL | 186.05 µL | 930.25 µL | 1.86 mL |
10 mM | 9.302 µL | 46.512 µL | 93.025 µL | 465.125 µL | 930.25 µL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Spectrum

Spectral properties
Correction Factor (260 nm) | 1.076 |
Correction Factor (280 nm) | 0.769 |
Extinction coefficient (cm -1 M -1) | 23430 |
Excitation (nm) | 495 |
Emission (nm) | 516 |
Quantum yield | 0.161 |
Product Family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield |
Fluo-8®, AM | 495 | 516 | 23430 | 0.161 |
Fluo-8L™, AM | 495 | 516 | 23430 | 0.161 |
Fluo-8FF™, AM | 495 | 516 | 23430 | 0.161 |
Fluo-4 AM *Ultrapure Grade* *CAS 273221-67-3* | 495 | 528 | 82000 | 0.161 |
Fluo-3, AM *CAS 121714-22-5* | 506 | 515 | 86,0001 | 0.151 |
Fluo-3, AM *UltraPure grade* *CAS 121714-22-5* | 506 | 515 | 86,0001 | 0.151 |
Fluo-3, AM *Bulk package* *CAS 121714-22-5* | 506 | 515 | 86,0001 | 0.151 |
Fluo-3FF, AM *UltraPure grade* *Cell permeant* | 506 | 515 | 86,0001 | 0.151 |
Fluo-5F, AM *Cell permeant* | 494 | 516 | - | - |
Show More (1) |
Images

Citations
Authors: Shiba, Kogiku and Inaba, Kazuo
Journal: Biomolecules (2023): 1594
Authors: Sato, Katsuya and Nakahara, Tasuku and Minami, Kazuyuki
Journal: Journal of Robotics and Mechatronics (2023): 1135--1142
Authors: Han, Shiqian and Wang, Qijun and Song, Yongfeng and Pang, Mao and Ren, Chunguang and Wang, Jing and Guan, Dongwei and Xu, Wei and Li, Fangyong and Wang, Fengchao and others,
Journal: iScience (2023)
Authors: Shiba, Kogiku and Inaba, Kazuo
Journal: International Journal of Molecular Sciences (2022): 1648
Authors: Isaeva, Elena and Bohovyk, Ruslan and Fedoriuk, Mykhailo and Shalygin, Alexey and Klemens, Christine A and Zietara, Adrian and Levchenko, Vladislav and Denton, Jerod S and Staruschenko, Alexander and Palygin, Oleg
Journal: British journal of pharmacology (2021)
Authors: Mazzarda, Flavia
Journal: (2021)
Authors: SATO, Katsuya and OOMORI, Daiki
Journal: Journal of Biomechanical Science and Engineering (2021): 21--00294
Authors: Mazzarda, Flavia and D'Elia, Annunziata and Massari, Roberto and De Ninno, Adele and Bertani, Francesca Romana and Businaro, Luca and Ziraldo, Gaia and Zorzi, Veronica and Nardin, Chiara and Peres, Chiara and others,
Journal: Lab on a Chip (2020): 3011--3023
Authors: Makino, Nanae and Sato, Nozomi and Takayama-Watanabe, Eriko and Watanabe, Akihiko
Journal: Reproduction (2020): 339--349
Authors: Bhandage, Amol K and Olivera, Gabriela C and Kanatani, Sachie and Thompson, Elizabeth and Lor{\'e}, Karin and Varas-Godoy, Manuel and Barragan, Antonio
Journal: Elife (2020): e60528
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