Details
The Protease Inhibitor Cocktail (100X, EDTA-free) contains an optimized mixture of six protease inhibitors prepared in DMSO as a ready-to-use 100X stock. The inhibitors (Aprotinin, AEBSF, Bestatin, E64, Leupeptin Hemisulfate, and Pepstatin A) in the cocktail provide a broad-spectrum protection against serine-proteases, cysteine-proteases, aspartic acid-proteases and aminopeptidases in mammalian cells and tissues, as well as in bacterial cells.
If EDTA is desired for inhibition of metalloproteases in an application, add EDTA (100X stock in water and provided in a separate vial) to the lysis buffer at a final working concentration of 5 mM.
To prepare 1X Protease Inhibitor Cocktail-containing lysis buffer:
1) Thaw the Protease Inhibitor Cocktail under room temperature or warm up in your palm.
2) Add an appropriate amount of the cocktail stock into lysis buffer directly (1 to 100 dilution, such as 0.1 ml cocktail solution into 9.9 ml lysis buffer, vortex immediately to help dissolve inhibitors. Lastly, add 0.1 ml EDTA.
| Component |
Protease Target |
Inhibitor Type |
100X Inhibitor Conc. |
| Aprotinin |
Serine |
Reversible |
0.08 mM |
| Pepstatin A |
Aspartic Acid |
Reversible |
1.0 mM |
| Bestatin |
Amino |
Reversible |
5.0 mM |
| AEBSF HCl |
Serine |
Irreversible |
100 mM |
| Leupeptin |
Serine/Cystine |
Reversible |
2.0 mM |
| E64D |
Cysteine |
Irresversible |
1.5 mM |
