Calbryte™ 630 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) | 1200 |
Molecular weight | 1234.84 |
Solvent | DMSO |
Excitation (nm) | 607 |
Emission (nm) | 624 |
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 1234.84 | Dissociation constant (Kd, nM) 1200 | Excitation (nm) 607 | Emission (nm) 624 |
Platform
Flow cytometer
Excitation | 640 nm laser |
Emission | 660/20 nm filter |
Instrument specification(s) | APC channel |
Fluorescence microscope
Excitation | Texas Red |
Emission | Texas Red |
Recommended plate | Black wall/clear bottom |
Fluorescence microplate reader
Excitation | 600 |
Emission | 640 |
Cutoff | 630 |
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™ 630 AM in anhydrous DMSO.
Note: When reconstituted in DMSO, Calbryte™ 630 AM is a clear, colorless solution.
PREPARATION OF WORKING SOLUTION
On the day of the experiment, either dissolve Calbryte™ 630 AM in DMSO or thaw an aliquot of the indicator stock solution to room temperature.
Prepare a 2 to 20 µM Calbryte™ 630 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™ 630 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™ 630 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™ 630 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 a Texas Red filter set or a fluorescence plate reader containing a programmable liquid handling system such as an FDSS, FLIPR, or FlexStation, at Ex/Em = 600/640 nm cutoff 630 nm.
Calculators
Common stock solution preparation
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 80.982 µL | 404.911 µL | 809.822 µL | 4.049 mL | 8.098 mL |
5 mM | 16.196 µL | 80.982 µL | 161.964 µL | 809.822 µL | 1.62 mL |
10 mM | 8.098 µL | 40.491 µL | 80.982 µL | 404.911 µL | 809.822 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™ 590 AM | 581 | 593 | - |
Calbryte™-520L AM | 493 | 515 | 0.751 |
Calbryte™-520XL AM | 493 | 515 | 0.751 |
Cal-630™ AM | 609 | 626 | 0.371 |
Images
Citations
Authors: Feng, Xiaogang and Andersson, Tilde and Gschwend, Julia and Fl{\"u}chter, Pascal and Berest, Ivan and Muff, Julian L and Carchidi, Daniele and Lechner, Antonie and de Tenorio, Jeshua C and Brander, Nina and others,
Journal: bioRxiv (2024): 2024--03
Authors: Acreman, Samuel and Ma, Jinfang and Denwood, Geoffrey and Gao, Rui and Tarasov, Andrei and Rorsman, Patrik and Zhang, Quan
Journal: iScience (2024)
Authors: Huang, Kate and Ashraf, Mishal and Rohani, Leili and Luo, Yinhan and Sacayanan, Ardin and Huang, Haojun and Haegert, Anne and Volik, Stanislav and Sar, Funda and LeBihan, St{\'e}phane and others,
Journal: bioRxiv (2023): 2023--03
Authors: Jin, Zihui and Zhao, Lingling and Chang, Yixin and Jin, Rongjia and Hu, Fangyu and Wu, Shuang and Xue, Zixuan and Ma, Yimeng and Chen, Chenglin and Zheng, Minghui and others,
Journal: Cell Death \& Disease (2023): 563
Authors: Zang, Shaolian and Yin, Xiaoqin and Li, Pin
Journal: Communications Biology (2023): 1297
Authors: DuBreuil, Daniel M and Lai, Xiaofan and Zhu, Kevin and Chahyadinata, Grace and Perner, Caroline and Chiang, Brenda and Battenberg, Ashley and Sokol, Caroline and Wainger, Brian
Journal: Molecular Pain (2022): 17448069221148351
Authors: Arslanova, Alia and Shafaattalab, Sanam and Ye, Kevin and Asghari, Parisa and Lin, Lisa and Kim, BaRun and Roston, Thomas M and Hove-Madsen, Leif and Van Petegem, Filip and Sanatani, Shubhayan and others,
Journal: Current Protocols (2021): e320
Authors: Liang, Qian and Wang, Yun and Lu, Yingsi and Zhu, Qingqing and Xie, Wenlin and Tang, Nannan and Huang, Lifen and An, Tailai and Zhang, Di and Yan, Anqi and others,
Journal: Cell death \& disease (2021): 1--18
Authors: Plante, Amber E and Meredith, Andrea L
Journal: Biophysical Journal (2019): 240a
Authors: Autry, Joseph M and Svensson, Bengt and Karim, Christine B and Perumbakkam, Sudeep and Chen, Zhenhui and Finno, Carrie J and Thomas, David D and Valberg, Stephanie J
Journal: Biophysical Journal (2019): 238a--239a
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
Calibration Protocol for Fluorescent Calcium Indicators
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
FAQ
Are there any calcium indicators that don't require probenecid (PBC)?
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?