Protease Assay Kit
Product Description: The Protease Assay Kit (sufficient for 250 assays) offers a simple non-radioactive
method for routine assays monitoring the presence of contaminating proteases in protein samples and
following the isolation of proteases. Casein is used as substrate in the assay kit, and its breakdown is
measured by a simple colorimetric protein-dye binding method. Degradation of casein by a protease causes a
decrease in absorbance at 595 nm. The assay kit can also be custom modified for determining cofactor
dependence, pH or temperature optima, and potential regulation by a protease activator or inhibitor. For
instance, including 5 mM or 5 mM Ca2+ in the assay buffer allows one to determine the activity of calpain I or
calpain II, respectively. A protease control (trypsin) is included in the kit, allowing users to perform a positive
control assay. A set of 10x concentrated buffers from pH 6 - 8 are provided for determining pH vs. activity
profiles of the proteases of interest. Assays can be performed in microplates and measured by a microplate
reader. Results can be obtained in less than 30 min. Assay conditions for a particular protease can be easily
optimized for maximal sensitivity.
Kit Components:
10x Buffer Set (pH 6 - 8): 1 ml each, store at -20ºC
Casein Solution (2 mg/ml): 1.5 ml, store at -20ºC
Protease Assay Solution: 25 ml, store at 4ºC
Product Description: The Protease Assay Kit (sufficient for 250 assays) offers a simple non-radioactive
method for routine assays monitoring the presence of contaminating proteases in protein samples and
following the isolation of proteases. Casein is used as substrate in the assay kit, and its breakdown is
measured by a simple colorimetric protein-dye binding method. Degradation of casein by a protease causes a
decrease in absorbance at 595 nm. The assay kit can also be custom modified for determining cofactor
dependence, pH or temperature optima, and potential regulation by a protease activator or inhibitor. For
instance, including 5 mM or 5 mM Ca2+ in the assay buffer allows one to determine the activity of calpain I or
calpain II, respectively. A protease control (trypsin) is included in the kit, allowing users to perform a positive
control assay. A set of 10x concentrated buffers from pH 6 - 8 are provided for determining pH vs. activity
profiles of the proteases of interest. Assays can be performed in microplates and measured by a microplate
reader. Results can be obtained in less than 30 min. Assay conditions for a particular protease can be easily
optimized for maximal sensitivity.
Kit Components:
10x Buffer Set (pH 6 - 8): 1 ml each, store at -20ºC
Casein Solution (2 mg/ml): 1.5 ml, store at -20ºC
Protease Assay Solution: 25 ml, store at 4ºC
Biomedical Research Service
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