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Description
Determine total protease activity in biological samples via fluorescent or colorimetric detection by measuring digestion of labeled casein substrate.
The Thermo Scientific Pierce Fluorescent Protease Assay Kit measures total protease activity in samples, providing a means to assess the progress of protease isolation procedures or quantify protease contamination in biological samples.
This kit includes fluorescein-labeled casein as a substrate for assessing protease activity in a sample by either fluorescence resonance energy transfer (FRET) with a standard fluorometer or fluorescence polarization (FP) with capable instrumentation. FTC-Casein is native casein that has been labeled using a large molar excess of fluorescein isothiocyanate (FITC). Fluorescence properties of this heavily-labeled, intact protein substrate change dramatically upon digestion by proteases, resulting in a measurable indication of proteolysis.
Features of the Fluorescent Protease Assay Kit:
- Casein substrate—measure activity of any protease that cleaves casein into peptide fragments
- Trypsin standard—quantify protease activity relative to trypsin, a universally accepted reference
- Sensitive—1000 times more sensitive than assays that use unmodified forms of casein
- Simple and fast—test tube and microplate protocols are completed in less than one hour
- Homogenous—addition steps only; no separation, transfer or stop steps required
- Customizable—easily adapt time, temperature and pH to optimize sensitivity for proteases of interest
Applications:
- Assess functional integrity and overall progress of protease purification
- Quantify protease contamination in protein samples
- Characterize the activity of unusual proteases compared to trypsin
- Evaluate buffer conditions for their affects on protease function and activity
Additional Specifications:
- Substrate: fluorescein-labeled casein (FITC-casein); available separately as Part No. 23267
- Basis of assay: change in fluorescence resonance (FRET) or fluorescence polarization (FP)
- Measurement: instrument with fluorescein excitation and emission filters (485/538nm)
Related Products
Pierce™ Colorimetric Protease Assay Kit (Cat. No. 23263)
FITC-Casein for Pierce™ Fluorescent Protease Assay Kit (Cat. No. 23267)
Specifications
Specifications
| Content And Storage | Sufficient For: 2000 microplate assays using an instrument fitted with a fluorescein filter set (ex/em 485 nm/538 nm) • FTC-Casein, lyophilized, 2.5 mg • TPCK Trypsin Standard, 50 mg • TBS Buffer Pack (makes 500 mL), 1 pack Store FTC-Casein and TPCK Trypsin at 4°C. |
| Assay | Protease Assay |
| Detection Method | Fluorescence |
| Immunoassay Kit Format | 96-well Plate |
| Label Type | Classic Dye |
| Product Line | Pierce |
| Product Type | Protein Activity Assay |
| Substrate | Casein |
| Substrate Properties | Protein-Based Substrate |
| Substrate Type | Protease Substrate |
| Show More |
Safety and Handling
- Pierce Fluorescent Protease Assay Kit
- Danger
- Serious eye damage/eye irritation Category 2
- Respiratory sensitiser Category 1
- Skin corrosion/irritation Category 2
- Skin sensitiser Category 1
- Specific target organ toxicity Category 3
- H315-Causes skin irritation.
- H317-May cause an allergic skin reaction.
- H319-Causes serious eye irritation.
- H334-May cause allergy or asthma symptoms or breathing difficulties if inhaled.
- H335-May cause respiratory irritation.
- P261-Avoid breathing dust/fume/gas/mist/vapours/spray.
- P272-Contaminated work clothing should not be allowed out of the workplace.
- P280-Wear protective gloves/protective clothing/eye protection/face protection.
- P284-[In case of inadequate ventilation] wear respiratory protection.
- P302+P352-IF ON SKIN: Wash with plenty of water/soap.
- P304+P340-IF INHALED: Remove person to fresh air and keep comfortable for breathing.
- P332+P313-If skin irritation occurs: Get medical advice/attention.
- P342+P311-If experiencing respiratory symptoms: Call a POISON CENTER/doctor/ .
- P363-Wash contaminated clothing before reuse.
- P501b-Dispose of contents/container in accordance with local/regional/national/international regulations.
- MIXTURE LIST-Contain: Sodium chloride, Tris (hydroxymethyl) aminomethane, Fluoresceinthiocarbamoyl-casein, Trypsin
For Research Use Only. Not for use in diagnostic procedures.
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