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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 Colorimetric 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 uses fully succinylated casein as a substrate for the assay. Hydrolysis of this substrate in the presence of protease results in the release of peptide fragments with free amino-terminal groups. These peptides are reacted with trinitrobenzene sulfonic acid (TNBSA), followed by measurement of the absorbance increase that results from the formation of yellow colored TNB-peptide adducts.
Features of the Colorimetric 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: succinylated casein (not available separately)
- Basis of assay: detection of primary amines resulting from proteolysis
- Detection reagent: trinitrobenzene sulfonic acid (TNBSA)
- Measurement: absorbance at 450nm (plate reader or spectrophotometer)
Related Products
Pierce™ Fluorescent Protease Assay Kit (Cat. No. 23266)
FITC-Casein for Pierce™ Fluorescent Protease Assay Kit (Cat. No. 23267)
Specifications
Specifications
| Content And Storage | Sufficient For: 250 microplate assays using a standard plate reader (absorbance at 450 nm) • Succinylated Casein, 5 x 10 mg • TNBSA Solution, 2 mL • TPCK Trypsin Standard, 50 mg • Borate Buffer Pack (makes 500 mL), 1 pack Store at 4°C. |
| Assay | Protease Assay |
| Detection Method | Absorbance |
| Immunoassay Kit Format | 96-well Plate |
| Product Line | Pierce |
| Product Type | Protein Activity Assay |
| Substrate | Casein |
| Substrate Properties | Protein-Based Substrate |
| Substrate Type | Protease Substrate |
| Sufficient For | 250 Microplate Assays |
| Show More |
Safety and Handling
- Pierce Colorimetric Protease Assay Kit
- Danger
- Flammable liquid Category 2
- Specific target organ toxicity Category 3
- Reproductive toxicity Category 1A
- Acute toxicity Category 3
- Skin corrosion/irritation Category 1 A
- Serious eye damage/eye irritation Category 1
- Skin sensitiser Category 1
- Specific target organ toxicity after single exposure Category 1
- Skin corrosion/irritation Category 2
- Serious eye damage/eye irritation Category 2
- Respiratory sensitiser Category 1
- H225-Highly flammable liquid and vapour.
- H335-May cause respiratory irritation.
- H360D-May damage the unborn child.
- H302-Harmful if swallowed.
- H311-Toxic in contact with skin.
- H314-Causes severe skin burns and eye damage.
- H317-May cause an allergic skin reaction.
- H331-Toxic if inhaled.
- H370-Causes damage to organs.
- H319-Causes serious eye irritation.
- H334-May cause allergy or asthma symptoms or breathing difficulties if inhaled.
- P280-Wear protective gloves/protective clothing/eye protection/face protection.
- P201-Obtain special instructions before use.
- P202-Do not handle until all safety precautions have been read and understood.
- P261-Avoid breathing dust/fume/gas/mist/vapours/spray.
- P302+P352-IF ON SKIN: Wash with plenty of water/.
- P301+P310-IF SWALLOWED: Immediately call a POISON CENTER/doctor /
- P303+P361+P353-IF ON SKIN (or hair): Take off immediately all contaminated clothing. Rinse skin with water/ shower.
- P305+P351+P338-IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing.
- P304+P340-IF INHALED: Remove person to fresh air and keep comfortable for breathing.
- P332+P313-If skin irritation occurs: Get medical advice/attention.
- P337+P313-If eye irritation persists: Get medical advice/attention.
- P308+P313-IF exposed or concerned: Get medical advice/attention.
- P370+P378-In case of fire: Use to extinguish.
- MIXTURE LIST-Contain: Methyl alcohol, Trypsin, Sodium tetraboratedecahydrate, Boric acid (H3BO3)
For Research Use Only. Not for use in diagnostic procedures.
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