Calbryte™ 590 AM
Price | |
Catalog Number | |
Availability | In stock |
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) | 1400 |
Molecular weight | 1218.77 |
Solvent | DMSO |
Excitation (nm) | 581 |
Emission (nm) | 593 |
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 |
Overview | SDSProtocol |
Molecular weight 1218.77 | Dissociation constant (Kd, nM) 1400 | Excitation (nm) 581 | Emission (nm) 593 |
Platform
Fluorescence microscope
Excitation | TRITC/Cy3 |
Emission | TRITC/Cy3 |
Recommended plate | Black wall/clear bottom |
Fluorescence microplate reader
Excitation | 540 |
Emission | 590 |
Cutoff | 570 |
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 Calbryte™ 590 AM in anhydrous DMSO.
Note: When reconstituted in DMSO, Calbryte™ 590 AM is a clear, colorless solution.
PREPARATION OF WORKING SOLUTION
On the day of the experiment, either dissolve Calbryte™ 590 AM in DMSO or thaw an aliquot of the indicator stock solution to room temperature.
Prepare a 2 to 20 µM Calbryte™ 590 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, Calbryte™ 590 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 Calbryte™ 590 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 Calbryte™ 590 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 TRITC/Cy3 filter set or a fluorescence plate reader containing a programmable liquid handling system such as an FDSS, FLIPR, or FlexStation, at Ex/Em = 540/590 nm cutoff 570 nm.
Calculators
Common stock solution preparation
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 82.05 µL | 410.25 µL | 820.499 µL | 4.102 mL | 8.205 mL |
5 mM | 16.41 µL | 82.05 µL | 164.1 µL | 820.499 µL | 1.641 mL |
10 mM | 8.205 µL | 41.025 µL | 82.05 µL | 410.25 µL | 820.499 µL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
/ | = | x | = |
Product Family
Name | Excitation (nm) | Emission (nm) | Quantum yield |
Calbryte™ 520 AM | 493 | 515 | 0.751 |
Calbryte™ 630 AM | 607 | 624 | - |
Calbryte™-520L AM | 493 | 515 | 0.751 |
Calbryte™-520XL AM | 493 | 515 | 0.751 |
Cal-590™ AM | 574 | 588 | 0.621 |
Images
Citations
Authors: Heider, Johanna and Stahl, Aaron and Sperlich, Denise and Hartmann, Sophia-Marie and Vogel, Sabrina and Breitmeyer, Ricarda and Templin, Markus and Volkmer, Hansj{\"u}rgen
Journal: BMC Neuroscience (2024): 1--20
Authors: Yu, Fang and Courjaret, Raphael and Assaf, Lama and Elmi, Asha and Hammad, Ayat and Fisher, Melanie and Terasaki, Mark and Machaca, Khaled
Journal: iScience (2024)
Authors: Liu, Hailin and Zhou, Lian and Wang, Xifeng and Zheng, Qingcui and Zhan, Fenfang and Zhou, Lanqian and Dong, Yao and Xiong, Yanhong and Yi, Pengcheng and Xu, Guohai and others,
Journal: Biochemical Pharmacology (2024): 116050
Authors: York, Nathaniel W and Patel, Sumit and Yan, Zihan and Tate, Abbie L and Frazier, Courtney and Remedi, Maria S and Nichols, Colin G
Journal: Biophysical Journal (2024): 263a
Authors: Miller, Zoey A and Mueller, Arielle and Kim, TaeBeom and Jolivert, Jennifer F and Ma, Ray Z and Muthuswami, Sahil and Park, April and McMahon, Derek B and Nead, Kevin T and Carey, Ryan M and others,
Journal: Cell Reports (2023)
Authors: Rybak-Wolf, Agnieszka and Wyler, Emanuel and Pentimalli, Tancredi Massimo and Legnini, Ivano and Oliveras Martinez, Anna and Gla{\v{z}}ar, Petar and Loewa, Anna and Kim, Seung Joon and Kaufer, Benedikt B and Woehler, Andrew and others,
Journal: Nature Microbiology (2023): 1--15
Authors: Emerson, James I and Ariel, Pablo and Shi, Wei and Conlon, Frank L
Journal: Journal of Cardiovascular Development and Disease (2023): 479
Authors: Luo, Zhiyong and Zhan, Zhipeng and Qin, Xiaowei and Pan, Weiqiang and Liang, Mincong and Li, Chuanrui and Weng, Shaoping and He, Jianguo and Guo, Changjun
Journal: Journal of Virology (2023): e00495--23
Authors: Smith, Holly A and Taylor, Colin W
Journal: Journal of Biological Chemistry (2023): 102871
Authors: Ved, Mansi
Journal: (2023)
References
Authors: Bailey S, Macardle PJ.
Journal: J Immunol Methods (2006): 220
Authors: Orlicky J, Sulova Z, Dovinova I, Fiala R, Zahradnikova A, Jr., Breier A.
Journal: Gen Physiol Biophys (2004): 357
Authors: Patel H, Porter RH, Palmer AM, Croucher MJ.
Journal: Br J Pharmacol (2003): 671
Authors: Loughrey CM, MacEachern KE, Cooper J, Smith GL.
Journal: Cell Calcium (2003): 1
Authors: Zhang Z, Kitching P.
Journal: J Virol Methods (2000): 187
Authors: Rockwell PL, Storey BT.
Journal: Mol Reprod Dev (2000): 335
Authors: Cantz T, Nies AT, Brom M, Hofmann AF, Keppler D.
Journal: Am J Physiol Gastrointest Liver Physiol (2000): G522
Authors: Walczysko P, Wagner E, Albrechtova JT.
Journal: Cell Calcium (2000): 23
Authors: Wohlfart B., undefined
Journal: Acta Physiol Scand (2000): 1
Authors: Rockwell PL, Storey BT.
Journal: Mol Reprod Dev (1999): 418
Application notes
Introducing Calbryte™ Series
A Comparison of Fluorescent Red Calcium Indicators for Detecting Intracellular Calcium Mobilization in CHO Cells
Calibration Protocol for Fluorescent Calcium Indicators
A New Robust No-Wash FLIPR Calcium Assay Kit for Screening GPCR and Calcium Channel Targets
FAQ
What are the differences between calcium ion indicators: Cal 520, Cal 520FF, and Cal 520N?
How do I make an AM ester stock solution?
How do you measure Ca2+ response with calcium indicators like Calbryte 520, potassium salt?
Can I intracellularly measure mitochondria calcium flux and changes in mitochondria membrane potential at the same time?