Screen Quest™ Luminometric Calcium Assay Kit
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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 |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12352200 |
Overview | ![]() ![]() |
Platform
Luminescence microplate reader
Recommended plate | Black wall/clear bottom |
Instrument specification(s) | Bottom read mode/Programmable liquid handling |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells by removing growth medium
- Add Coelenterazine-loading solution (100 µL/well for 96-well plate or 25 µL/well for 384-well plate)
- Incubate at room temperature for 3-4 hours
- Monitor aequorin luminescence intensity
Important notes
Thaw all the kit components at room temperature before starting the experiment.
PREPARATION OF STOCK SOLUTION
1. Coelenterazine analog:
Add 250 µL of 100% ETOH (Component B) into the vial of Coelenterazine analog (Component A), and mix them well. Note: 25 µL of reconstituted coelenterazine analog is enough for one plate. Unused coelenterazine analog stock solution can be stored at < -20 oC for more than one month if the tubes are sealed tightly. Protect from light and avoid repeated freeze-thaw cycles.
2. Assay Buffer (1X):
a) For Cat. # 36305 (10 plates kit), ready to use 1X Assay Buffer (Component C).
b)For Cat. # 36306 (100 plates kit), make 1X assay buffer by diluting 10 mL of 10X Assay Buffer (Component C) into 90 mL of HHBS buffer (not included in the kit), and mix them well. Note: 10 mL of 1X assay buffer is enough for one plate. Store unused 1X assay buffer at 4 oC.
PREPARATION OF WORKING SOLUTION
Coelenterazine-loading solution:
Add 25 µL of ETOH reconstituted coelenterazine analog (Prepartion Of Stock Solutions) into 10 mL of 1X assay buffer (Preparation Of Stock Solutions), and mix them well. Note: This working solution is stable for at least 2 hours at room temperature, protected from light.
For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html
SAMPLE EXPERIMENTAL PROTOCOL
- Remove the growth medium from the cell plates. Note: It is important to remove the growth medium in order to minimize compound interference with serum or culture media. Note: Alternatively, grow the cells in growth medium with 0.5-1% FBS to avoid medium removal step. In this case, 2X Coelenterazine-loading solution in 1X buffer is needed.
- Add 100 µL/well (96-well plate) or 25 µL/well (384-well plate) Coelenterazine-loading solution into the cell plates.
- Incubate the Coelenterazine-loading plates at room temperature for 3-4 hours, protected from light.
- Prepare the compound plates with HHBS or the desired buffer.
- Monitor the aequorin luminescence intensity by using the photon detection system that has an enclosed chamber containing a photomultiplier. The instrument must completely exclude outside light.
Images
![ATP Dose Response on CHO-aeq cells. CHO cells stably transfected with apoaequrin were seeded overnight at 50,000 cells/100 µL/well in a Costar white wall/clear bottom 96-well plate. The growth medium was removed and the cells were incubated with 100 µL of dye-loading solution using the Screen Quest™ Coelenterazine Calcium Assay Kit for 3 hours at room temperature and protected from light. ATP (25 µL/well) was added by NOVOstar (BMG Labtech) to achieve the final indicated concentrations. The EC50 of ATP is about 0.8 µM.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fscreen-quest-luminometric-calcium-assay-kit%2Fgraph-for-screen-quest-luminometric-calcium-assay-kit_sLetD.webp&w=3840&q=75)
Citations
Authors: Ma, Yi and Zhao, Yichen and Berkowitz, Gerald A
Journal: Journal of Experimental Botany (2017)
Authors: Harikumar, Kaleeckal G and Yan, Yan and Xu, Ting-Hai and Melcher, Karsten and Xu, H Eric and Miller, Laurence J
Journal: (2017)
Authors: Kim, Donghun and Simo, Ladislav and Park, Yoonseong
Journal: Journal of Experimental Biology (2014): 3656--3663
References
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Journal: J Mol Cell Cardiol (2006): 980
Authors: Inouye S, Sasaki S.
Journal: FEBS Lett (2006): 1977
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Journal: Nat Biotechnol (2006): 326
Authors: Inouye S., undefined
Journal: Protein Expr Purif. (2006)
Authors: Otto-Duessel M, Khankaldyyan V, Gonzalez-Gomez I, Jensen MC, Laug WE, Rosol M.
Journal: Mol Imaging (2006): 57
Authors: Liu ZJ, Stepanyuk GA, Vysotski ES, Lee J, Markova SV, Malikova NP, Wang BC.
Journal: Proc Natl Acad Sci U S A (2006): 2570
Authors: Pfleger KD, Dromey JR, Dalrymple MB, Lim EM, Thomas WG, Eidne KA.
Journal: Cell Signal (2006): 1664
Authors: Teranishi K., undefined
Journal: Bioorg Chem. (2006)
Authors: Tsuji FI., undefined
Journal: Biochem Biophys Res Commun (2005): 250
Application notes
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