WB
Product Information
Product Usage Information
1. Briefly vortex the Protease Inhibitor Cocktail (100X) before use.
2. Just prior to lysing cells, dilute the cocktail 1:100 in desired lysis buffer to obtain a 1X working concentration.
Solutions and Reagents
The Protease Inhibitor Cocktail (100X) is composed of a proprietary mix of AEBSF, Aprotinin, Bestatin, E64, Leupeptin, and Pepstatin A to promote broad spectrum protection against endogenous proteases. The cocktail does not contain EDTA (a metalloprotease inhibitor) which can be incompatible with some downstream applications (i.e. protein assays, 2D electrophoresis, etc.). If EDTA is desired as a protease inhibitor it can be added to the cell lysis buffer at a final working concentration of 5 mM.
Storage
Product Description
When diluted in lysis buffer to a final concentration of 1X the Protease Inhibitor Cocktail prevents protein degradation by endogenous proteases present in whole cell extract. The 100X Protease Inhibitor Cocktail is a clear, colorless liquid.
1X Protease Inhibitor Cocktail:
1mM AEBSF
800nM Aprotinin
50μM Bestatin
15μM E-64
20μM Leupeptin
10μM Pepstatin A
Background
In order to study specific target proteins of interest protease-mediated degradation during the generation of protein lysates is to be avoided. A loss of normal cellular control occurs during cell lysis, and endogenous proteases within the cell extract are free to degrade proteins in an uncontrolled manner. The addition of protease inhibitors to the cell lysis buffer aids in the preservation of target proteins in the cell extract.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Applications Key
WB: Western Blotting
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
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