Cleaning protocols for crystallization robots: preventing protease contamination
The protease in the commonly used commercial low‐foam enzyme cleaner Zymit cannot be completely blocked by EDTA, a widely used inhibitor of metalloproteases, at concentrations of up to 5 mM. Severe protein degradation was observed in crystallization drops after EDTA‐containing wash steps unless resi...
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Veröffentlicht in: | Acta crystallographica. Section F, Structural biology communications Structural biology communications, 2015-01, Vol.71 (1), p.100-102 |
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Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | The protease in the commonly used commercial low‐foam enzyme cleaner Zymit cannot be completely blocked by EDTA, a widely used inhibitor of metalloproteases, at concentrations of up to 5 mM. Severe protein degradation was observed in crystallization drops after EDTA‐containing wash steps unless residual Zymit protease was removed with NaOH at a concentration of at least 0.1 M. Wash steps with 0.1% SDS were also ineffective in completely removing the remaining Zymit activity. Protocols including wash steps with at least 0.1 M NaOH, as for example specified in the original ZENM protocol, are recommended to completely deactivate Zymit protease activity. |
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ISSN: | 2053-230X 2053-230X |
DOI: | 10.1107/S2053230X14026053 |