EGTA tetrasodium salt *CAS 13368-13-3*
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Molecular weight | 468.28 |
Solvent | Water |
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 |
EGTA tetrasodium salt *10 mM aqueous solution* |
Overview | SDSProtocol |
CAS 13368-13-3 | Molecular weight 468.28 |
Example protocol
AT A GLANCE
Important
Expiration date is 12 months from the date of receipt.
SAMPLE EXPERIMENTAL PROTOCOL
Sample Protocol for Calcium Measurement:
Titrate the concentration of free Ca2+ in solution by mixing different amounts of K2EGTA and CaEGTA. The reactions of these solutions with Fluo-3, Fluo 4, Fluo-8, Cal-520, or Calbryte 520 dye should be at room temperature, pH 7.2 and 100 mM KCl. Under these conditions, the Kd for EGTA is 150 nM. Measure the Fluo-3, Fluo 4, Fluo-8, Cal-520, or Calbryte 520 fluorescence intensity with a fluorescence microplate reader at Ex/Em = 490/525 nm.
- Mix the relative volumes of K2EGTA (Cat.# 21008) and CaEGTA according to the following table.
Sample # |
Volume K2EGTA, uL |
Volume CaEGTA, uL |
Calculated free Ca2+, uM |
RFU |
zero (blank) |
1000 |
0 |
0 |
0 |
1 |
900 |
100 | ||
2 |
800 |
200 | ||
3 |
700 |
300 | ||
4 |
600 |
400 | ||
5 |
500 |
500 | ||
6 |
400 |
600 | ||
7 |
300 |
700 | ||
8 |
200 |
800 | ||
9 |
100 |
900 |
- Calculate the concentration of free Ca2+ in each solution using the following formula:
[Ca2+]free = KdEGTA x {[CaEGTA]/[K2EGTA]} (Note: the Kd of EGTA is 150 nM).
- Add 1 uL of 1 mM Fluo-3, Fluo 4, Fluo-8, Cal-520, or Calbryte 520 into each solution including the blank.
- Read the fluorescence intensity of each solution with a fluorescent microplate reader at Ex/Em = 490/525 nm.
Calculators
Common stock solution preparation
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 213.547 µL | 1.068 mL | 2.135 mL | 10.677 mL | 21.355 mL |
5 mM | 42.709 µL | 213.547 µL | 427.095 µL | 2.135 mL | 4.271 mL |
10 mM | 21.355 µL | 106.774 µL | 213.547 µL | 1.068 mL | 2.135 mL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Citations
Authors: Viswanathan, Meera C and Schmidt, William and Rynkiewicz, Michael J and Agarwal, Karuna and Gao, Jian and Katz, Joseph and Lehman, William and Cammarato, Anthony
Journal: Cell Reports (2017): 2612--2625
Authors: Pecze, László and Blum, Walter and Schwaller, Beat
Journal: Journal of Biological Chemistry (2015): 28214--28230
References
Authors: Ellis-Davies GC, Barsotti RJ.
Journal: Cell Calcium (2006): 75
Authors: Fisher AE, Hague TA, Clarke CL, Naughton DP.
Journal: Biochem Biophys Res Commun (2004): 163
Authors: Schuhmeier RP, Dietze B, Ursu D, Lehmann-Horn F, Melzer W.
Journal: Biophys J (2003): 1065
Authors: Belyavskaya NA., undefined
Journal: J Gravit Physiol (2001): P33
Authors: Shivakumar K, Kumaran C.
Journal: J Mol Cell Cardiol (2001): 373
Authors: Boullerne AI, Nedelkoska L, Benjamins JA.
Journal: J Neurosci Res (2001): 124
Authors: Levin RM, Leggett R, Whitbeck C, Horan P.
Journal: Pharmacology (1999): 113
Authors: Pape PC, Carrier N.
Journal: J Gen Physiol (1998): 161
Authors: Pape PC, Jong DS, Ch and ler WK., undefined
Journal: J Gen Physiol (1998): 263
Authors: Struk A, Szucs G, Kemmer H, Melzer W.
Journal: Cell Calcium (1998): 23
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
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