Fetal calf sera can distort cell-based luminescent proteasome assays through heat-resistant chymotrypsin-like activity
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. Usi...
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| Main Authors: | , , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
2015
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| In: |
Analytical biochemistry
Year: 2014, Volume: 471, Pages: 23-25 |
| ISSN: | 1096-0309 |
| DOI: | 10.1016/j.ab.2014.10.019 |
| Online Access: | Verlag, Volltext: http://dx.doi.org/10.1016/j.ab.2014.10.019 Verlag, Volltext: http://www.sciencedirect.com/science/article/pii/S0003269714005090 
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| Author Notes: | Susan Dettmer, Dirk Theile, Anja Seckinger, Jürgen Burhenne, Johanna Weiss |
| Summary: | 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. |
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| Item Description: | Published online: 4 November 2014 Gesehen am 17.07.2017 |
| Physical Description: | Online Resource |
| ISSN: | 1096-0309 |
| DOI: | 10.1016/j.ab.2014.10.019 |