Quin-2, AM *CAS 83104-85-2*
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Dissociation constant (Kd, nM) | 60 |
Molecular weight | 829.76 |
Solvent | DMSO |
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 | SDSProtocol |
CAS 83104-85-2 | Molecular weight 829.76 | Dissociation constant (Kd, nM) 60 |
Platform
Fluorescence microplate reader
Excitation | 340 |
Emission | 495 |
Cutoff | 475 |
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 Quin-2 AM in high-quality, anhydrous DMSO.
PREPARATION OF WORKING SOLUTION
On the day of the experiment, either dissolve Quin-2 AM in DMSO or thaw an aliquot of the indicator stock solution to room temperature.
Prepare a 2 to 20 µM Quin-2 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, Quin-2 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 Quin-2 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 Quin-2 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 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 a fluorescence plate reader containing a programmable liquid handling system such as a FlexStation, at Ex/Em = 340/495 cutoff 475 nm.
Calculators
Common stock solution preparation
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 120.517 µL | 602.584 µL | 1.205 mL | 6.026 mL | 12.052 mL |
5 mM | 24.103 µL | 120.517 µL | 241.034 µL | 1.205 mL | 2.41 mL |
10 mM | 12.052 µL | 60.258 µL | 120.517 µL | 602.584 µL | 1.205 mL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
/ | = | x | = |
Product Family
Name | Excitation (nm) | Emission (nm) |
Fura-2, AM *CAS 108964-32-5* | 336 | 505 |
Fura-2, AM *UltraPure Grade* *CAS 108964-32-5* | 336 | 505 |
Rhod-2, AM *CAS#: 145037-81-6* | 553 | 577 |
Rhod-2, AM *UltraPure Grade* *CAS#: 145037-81-6* | 553 | 577 |
References
Authors: Goko H, Matsuoka A.
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Journal: Cell Calcium (1994): 7
Authors: Rao GH, Gerrard JM, Murthy M, White JG.
Journal: Biochem Med Metab Biol (1993): 322
Authors: Lakowicz JR, Szmacinski H, Nowaczyk K, Johnson ML.
Journal: Cell Calcium (1992): 131
Authors: Carpenter-Deyo L, Duimstra JR, Hedstrom O, Reed DJ.
Journal: J Pharmacol Exp Ther (1991): 739
Authors: Carpenter-Deyo L, Reed DJ.
Journal: J Pharmacol Exp Ther (1991): 747
Authors: Rao GH, White JG.
Journal: Prostaglandins Leukot Essent Fatty Acids (1990): 141
Authors: Blanco P, Martinez-Serrano A, Bogonez E, Satrustegui J.
Journal: Cell Calcium (1990): 25
Authors: Mazorow DL, Millar DB.
Journal: Anal Biochem (1990): 28
Application notes
A New Robust No-Wash FLIPR Calcium Assay Kit for Screening GPCR and Calcium Channel Targets
A Novel NO Wash Probeniceid-Free Calcium Assay for Functional Analysis of GPCR and Calcium Channel Targets
Evaluation of FLIPR Calcium Assays for Screening GPCR and Calcium Channel Targets
Fluorescent Dye AM Esters
FAQ
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