PIERCE 23263

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

Contents & Storage:

• Sufficient For: 250 microplate assays using a standard plate reader (absorbance at 450 nm)
• Succinylated Casein, 5 x 10mg
• TNBSA Solution, 2mL
• TPCK Trypsin Standard, 50mg
• Borate Buffer Pack (makes 500mL), 1 pack