OG488 BAPTA-1, AM [equivalent to Oregon Green® 488 BAPTA-1, AM] *Cell permeant*
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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) | 170 |
Molecular weight | 1258.07 |
Solvent | DMSO |
Excitation (nm) | 493 |
Emission (nm) | 522 |
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 |
Molecular weight 1258.07 | Dissociation constant (Kd, nM) 170 | Excitation (nm) 493 | Emission (nm) 522 |
Platform
Flow cytometer
Excitation | 488 nm laser |
Emission | 530/30 nm filter |
Instrument specification(s) | FITC channel |
Fluorescence microscope
Excitation | FITC filter set |
Emission | FITC filter set |
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 OG488 BAPTA-1 AM in high-quality, anhydrous DMSO.
PREPARATION OF WORKING SOLUTION
On the day of the experiment, either dissolve OG488 BAPTA-1 AM in DMSO or thaw an aliquot of the indicator stock solution to room temperature.
Prepare a 2 to 20 µM OG488 BAPTA-1 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, OG488 BAPTA-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 OG488 BAPTA-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 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 OG488 BAPTA-1 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 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 Ex/Em = 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 | 79.487 µL | 397.434 µL | 794.868 µL | 3.974 mL | 7.949 mL |
5 mM | 15.897 µL | 79.487 µL | 158.974 µL | 794.868 µL | 1.59 mL |
10 mM | 7.949 µL | 39.743 µL | 79.487 µL | 397.434 µL | 794.868 µL |
Molarity calculator
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Images
Citations
Authors: Birkner, Antje
Journal: (2019)
References
Authors: Schnell C, Negm M, Driehaus J, Scheller A, Hulsmann S.
Journal: Respir Physiol Neurobiol (2016): 18
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Journal: J Neurophysiol (2013): 243
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Journal: J Comp Physiol A Neuroethol Sens Neural Behav Physiol (2012): 477
Authors: Langer D, Helmchen F.
Journal: Pflugers Arch (2012): 339
Authors: Takata N, Mishima T, Hisatsune C, Nagai T, Ebisui E, Mikoshiba K, Hirase H.
Journal: J Neurosci (2011): 18155
Authors: Lattarulo C, Thyssen D, Kuchibholta KV, Hyman BT, Bacskaiq BJ.
Journal: Methods Mol Biol (2011): 377
Authors: Wotzlaw C, Bernardini A, Berchner-Pfannschmidt U, Papkovsky D, Acker H, F and rey J., undefined
Journal: Am J Physiol Cell Physiol (2011): C266
Authors: Bouhours B, Trigo FF, Marty A.
Journal: J Neurosci (2011): 5804
Authors: Misfeldt MW, Aalkjaer C, Simonsen U, Bek T.
Journal: Exp Eye Res (2010): 69
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
How do I make an AM ester stock solution?
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