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MeO-Succ-Arg-Pro-Tyr-AMC
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
MeO-Succ-Arg-Pro-Tyr-AMC is provided as a lyophilized powder. To prepare a 10 mM stock solution, dissolve 1 mg of the compound in 122 µL of DMSO.
PREPARATION OF WORKING SOLUTION
Prepare a substrate working solution at a final concentration of 20–50 µM using the reaction buffer (100 mM Tris-HCl, pH 8.0, containing 0.4 M NaCl and 0.1% Triton X-100).
Note: Prepare the MeO-Succ-Arg-Pro-Tyr-AMC working solution immediately before each experiment, and protect it from light.
SAMPLE EXPERIMENTAL PROTOCOL
Well | Volume |
STD1-STD7 | 50 µL AMC serial dilutions (0.078 µM-10 µM) |
BL | 50 µL reaction buffer |
Test Sample | 50 µL |
Sample Buffer Control | 50 µL of the buffer that the test sample is in |
Prepare the AMC standards (STD1–STD7), blank controls (BL), and test samples according to the layout specified in the table above.
Add 50 μL of Working Solution to each well designated for Test Samples and Sample Buffer Control. For STD1-STD7 and BL wells, add 50 μL reaction buffer.
Incubate the samples at room temperature for 30–60 minutes, protected from light. Or, monitor the reaction in real time using kinetic mode.
Monitor the fluorescence at Ex/Em= 360/470 nm (cutoff=435 nm).
Calculators
Common stock solution preparation
| 0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
| 1 mM | 121.984 µL | 609.92 µL | 1.22 mL | 6.099 mL | 12.198 mL |
| 5 mM | 24.397 µL | 121.984 µL | 243.968 µL | 1.22 mL | 2.44 mL |
| 10 mM | 12.198 µL | 60.992 µL | 121.984 µL | 609.92 µL | 1.22 mL |
Molarity calculator
| Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Spectrum
Citations
Authors: Fukagawa, Satoko and Takahashi, Ayami and Sayama, Keimon and Mori, Shinobu and Murase, Takatoshi
Journal: International Journal of Cosmetic Science (2020)
References
Authors: Bodenreider C, Beer D, Keller TH, Sonntag S, Wen D, Yap L, Yau YH, Shochat SG, Huang D, Zhou T, Caflisch A, Su XC, Ozawa K, Otting G, Vasudevan SG, Lescar J, Lim SP.
Journal: Anal Biochem (2009): 195
Authors: Wang X, Guo J, Wang J, Li S, Sun L, Lu J.
Journal: Sheng Wu Gong Cheng Xue Bao (2009): 701
Authors: Tsai MT, Cheng YH, Liu YN, Liao NC, Lu WW, Kung SH.
Journal: Antimicrob Agents Chemother (2009): 748
Authors: Mizukami S, Takikawa R, Sugihara F, Shirakawa M, Kikuchi K.
Journal: Angew Chem Int Ed Engl (2009): 3641
Authors: Zhang L, Xu Q, Xing D, Gao C, Xiong H.
Journal: Plant Physiol (2009): 1773
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