When extracting proteins from cells or tissues, endogenous and exogenous proteases rapidly degrade the proteins. Protein hydrolysis greatly reduces the quantity and activity of protein samples, increasing the difficulty of data analysis. To ensure the success of downstream research, adding a mixture of protease inhibitors and highly specific protease inhibitors during the cell lysis process can ensure protein integrity during protein extraction and preparation.
Classification of proteolytic enzymes:
Proteases are usually divided into four categories, with different types and abundances of proteases derived from different materials.
Serine proteases: including trypsin, chymotrypsin, and elastase
Cysteine proteases: including papain, calpain, and lysosomal tissue protease
Aspartate proteases: including pepsin and chymotrypsin
• Metalloproteinases: including thermolysins and carboxypeptidases
Protease inhibitors can inhibit the hydrolysis of various proteases, prevent protein degradation, and ensure protein yield and integrity. EDTA free in the mixture of protease inhibitors is specifically used for metal chelation affinity purification (IMAC) such as His tags.
choice
In most daily experiments, researchers choose the following protease inhibition mixture products:
Roche pioneered the concept of cocktail optimized formula (formula confidentiality), replacing single component inhibitors to provide comprehensive protein protection. The invention of EASypack aluminum foil packaging is convenient to use, further avoiding researchers from contacting toxic reagents and ensuring operational safety.
Sigma: Provides a comprehensive range of single component inhibitors, as well as inhibitor mixtures such as concentrates and tablets. The formula is fully disclosed, making it easy for users to evaluate; The working fluid is formulated at a ratio of 1:100 with extremely high cost-effectiveness; Specially provide a mixture of non animal derived protease inhibitors and specialized inhibitors for mass spectrometry samples.
Millipore Calbiochem: a leading traditional inhibitor brand with high cost-effectiveness.
