Casein, FITC-conjugated
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Molecular weight | N/A |
Solvent | Water |
Correction Factor (280 nm) | 0.35 |
Extinction coefficient (cm -1 M -1) | 73000 |
Excitation (nm) | 491 |
Emission (nm) | 516 |
Quantum yield | 0.92 |
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 N/A | Correction Factor (280 nm) 0.35 | Extinction coefficient (cm -1 M -1) 73000 | Excitation (nm) 491 | Emission (nm) 516 | Quantum yield 0.92 |
Example protocol
AT A GLANCE
Chemical Properties of Casein
Appearance: Light yellow powder
Excitation/Emission: 494/521 nm
Solvent: Water
PREPARATION OF STOCK SOLUTION
Casein, FITC-conjugated stock solution:
Make a 5 - 10 mg/mL Casein, FITC-conjugated stock solution in PBS buffer.
PREPARATION OF WORKING SOLUTION
Casein FITC-conjugated working solution (2X):
Dilute the FITC-conjugated stock solution into 50 - 100 mM Tris buffer (pH 7.4) at 100 - 400 μg/mL. The 2X Assay working solution is designed for detecting the activity of chymotrypsin, trypsin, thermolysin, proteinase K, protease XIV, and human leukocyte elastase. For other proteases, please refer to Table 1 for the appropriate assay buffer formula. The optimum concentration of the assay working solution should be determined experimentally for individual proteases.
SAMPLE EXPERIMENTAL PROTOCOL
- Mix equal volume of the trypsin standards or samples with 2X Assay working solution.
- Monitor the fluorescence increase at Ex/Em = 490/525 nm.
a. For kinetic reading: Immediately start measuring fluorescence intensity continuously and record data every 5 minutes for 30 minutes.
b. For end-point reading: Incubate the reaction at a desired temperature for 30 to 60 minutes, protected from light. Then measure the fluorescence intensity.
Table 1. Appropriate assay buffer formula for Assay working solution.
Protease | 1X Assay Buffer |
Cathepsin D | 20 mM Sodium Citrate, pH 3.0 |
Papain | 20 mM sodium acetate, 20 mM cysteine, 2 mM EDTA, pH 6.5 |
PAE | 20 mM sodium phosphate, pH 8.0 |
Pepsin | 10 mM HCl, pH 2.0 |
Porcine pancreas elastase | 10 mM Tris-HCl, pH 8.8 |
Subtilisin | 20 mM potassium phosphate buffer, pH 7.6, 150 mM NaCl |
Spectrum
Spectral properties
Correction Factor (280 nm) | 0.35 |
Extinction coefficient (cm -1 M -1) | 73000 |
Excitation (nm) | 491 |
Emission (nm) | 516 |
Quantum yield | 0.92 |
Product Family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Correction Factor (260 nm) | Correction Factor (280 nm) |
Casein, TAMRA-conjugated | 552 | 578 | 90000 | 0.32 | 0.178 |
Images
Citations
Authors: Tatsumi, Kazuya and Hsu, Chi Hsuan and Suzuki, Atsushi and Nakabe, Kazuyoshi
Journal: Heat and Mass Transfer (2018): 2607--2616
Authors: Tatsumi, Kazuya and Hsu, Chi Hsuan and Suzuki, Atsushi and Nakabe, Kazuyoshi
Journal: Heat and Mass Transfer (2017): 1--10
Authors: Tatsumi, K and Hsu, CH and Suzuki, A and Nakabe, K
Journal: (2016): 032097
Authors: Tatsumi, Kazuya and Tozaki, Akihisa and Nakabe, Kazuyoshi
Journal: (2011): T10167--T10167
References
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Journal: J Dairy Sci (2005): 3392
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Journal: J Biol Chem (2003): 11489
Authors: Lee EH, Kim CS, Cho JB, Ahn KJ, Kim KH.
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Journal: Biochemistry (2001): 9317
Authors: Scott ME, Dossani ZY, S and kvist M., undefined
Journal: Proc Natl Acad Sci U S A (2001): 13978
Authors: Gray CW, Ward RV, Karran E, Turconi S, Rowles A, Viglienghi D, Southan C, Barton A, Fantom KG, West A, Savopoulos J, Hassan NJ, Clinkenbeard H, Hanning C, Amegadzie B, Davis JB, Dingwall C, Livi GP, Creasy CL.
Journal: Eur J Biochem (2000): 5699
Authors: Fajardo-Lira C, Oria M, Hayes KD, Nielsen SS.
Journal: J Dairy Sci (2000): 2190
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