SunRed™ Phosphate
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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 |
Molecular weight | 422.20 |
Solvent | Water |
Excitation (nm) | 653 |
Emission (nm) | 661 |
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 | 12171501 |
Overview | SDSProtocol |
Molecular weight 422.20 | Excitation (nm) 653 | Emission (nm) 661 |
Platform
Fluorescence microplate reader
Excitation | 620 nm |
Emission | 660 nm |
Cutoff | 640 nm |
Recommended plate | Solid black |
Example protocol
AT A GLANCE
Protocol summary
- Prepare and add 10 - 50 µM SunRed™ Phosphate in Tris buffer (50 µL)
- Add alkaline phosphatase standards and/or test samples (50 µL)
- Incubate at room temperature or 37°C for 30 to 120 minutes
- Monitor fluorescence intensity at Ex/Em = 620/660 nm (cut off 640 nm)
Important notes
The following is the recommended protocol for alkaline phosphatase assay in solution. The protocol only provides a guideline, should be modified according to the specific needs.
PREPARATION OF STOCK SOLUTION
1. SunRed™ Phosphate stock solution:
Prepare a 2 to 10 mM stock solution of SunRed™ Phosphate in sterile water. Note: The stock solution should be used promptly. Note: Do not use DMSO, ETOH or METH to make stock solution since it significantly increases assay background. Note: Protect from light.
PREPARATION OF WORKING SOLUTION
SunRed™ Phosphate working solution (2X):
On the day of the experiment, either dissolve SunRed™ Phosphate solid in sterile H2O or thaw an aliquot of the SunRed™ Phosphate stock solution at room temperature. Prepare a 2X working solution of 10 to 50 µM in 100 mM Tris buffer or buffer of your choice, pH 8 to 9. SunRed™ Phosphate final concentration of 5 to 25 μM is recommended for measuring alkaline phosphatase activity in solution.
SAMPLE EXPERIMENTAL PROTOCOL
- Add 50 µL of 2X SunRed™ Phosphate working solution into each well of the alkaline phosphatase standard, blank control, and test samples to make the total alkaline phosphatase assay volume of 100 µL/well. For a 384-well plate, add 25 µL of sample and 25 µL of 2X SunRed™ Phosphate working solution into each well.
- Incubate the reaction for 30 to 120 minutes at the desired temperature, protected from light.
- Monitor the fluorescence increase at Ex/Em = 620/660 nm (cut off at 640 nm) with a fluorescence plate reader.
Calculators
Common stock solution preparation
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 236.855 µL | 1.184 mL | 2.369 mL | 11.843 mL | 23.685 mL |
5 mM | 47.371 µL | 236.855 µL | 473.709 µL | 2.369 mL | 4.737 mL |
10 mM | 23.685 µL | 118.427 µL | 236.855 µL | 1.184 mL | 2.369 mL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
/ | = | x | = |
Product Family
Name | Excitation (nm) | Emission (nm) | Correction Factor (280 nm) |
SunRed™ SE | 592 | 609 | 0.366 |
SunRed™ Acetate | 653 | 661 | - |
Images
Citations
Authors: Schmalenberg, Michael Reiner
Journal: (2017)
References
Authors: Marchetti S, Onori G, Cametti C.
Journal: J Phys Chem B (2005): 3676
Authors: Martin K, Hart C, Liu J, Leung WY, Patton WF.
Journal: Proteomics (2003): 1215
Authors: Top KP, Hatleberg G, Berggren KN, Ryan D, Kemper C, Haugl and RP, Patton WF.
Journal: Electrophoresis (2001): 896
Authors: Leira F, Vieites JM, Vieytes MR, Botana LM.
Journal: Toxicon (2000): 1833
Authors: Vazquez-Contreras E, de Gomez-Puyou MT, Dreyfus G.
Journal: Protein Expr Purif (1996): 55
Authors: Valerio M, Diolez P, Haraux F.
Journal: Eur J Biochem (1993): 565
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