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Fetal calf sera can distort cell-based luminescent proteasome assays through heat-resistant chymotrypsin-like activity
Authors:Susan Dettmer,Dirk Theile,Anja Seckinger,Jü  rgen Burhenne,Johanna Weiss
Affiliation:1. Department of Clinical Pharmacology and Pharmacoepidemiology, Heidelberg University Hospital, D-69120 Heidelberg, Germany;2. Department of Internal Medicine V, Oncology, Hematology, and Rheumatology, Heidelberg University Hospital, D-69120 Heidelberg, Germany
Abstract:
Luminescence-based proteasome activity assays use specific substrates that are supposed to be cleaved by cellular proteasome activity leading to luciferase substrates. Usually, control wells containing cell culture medium supplemented with antibiotics and fetal calf serum are used as background. Using the Proteasome-Glo chymotrypsin-like cell-based assay from Promega, we show here that fetal calf sera from different manufacturers contain heat-resistant, bortezomib-inhibitable, chymotrypsin-like activities that can interfere with proteasome activity assays. These data strongly recommend the use of pure phosphate-buffered saline (PBS) or serum-free medium during proteasome activity assays to diminish background luminescence and, thus, to obtain reliable results.
Keywords:Proteasome activity   Proteasome-Glo chymotrypsin-like cell-based assays   Fetal calf serum   Heat resistance   Bortezomib   Carfilzomib
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