TY - JOUR
T1 - Cleaning protocols for crystallization robots: Preventing protease contamination
AU - Naschberger, Andreas
AU - Fürnrohr, Barbara G.
AU - Dunzendorfer-Matt, Theresia
AU - Bonagura, Christopher A.
AU - Wright, David
AU - Scheffzek, Klaus
AU - Rupp, Bernhard
N1 - Generated from Scopus record by KAUST IRTS on 2023-02-15
PY - 2015/1/1
Y1 - 2015/1/1
N2 - 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 5mM. 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 0.1M. Wash steps with 0.1% SDS were also ineffective in completely removing the remaining Zymit activity. Protocols including wash steps with at least 0.1M NaOH, as for example specified in the original ZENM protocol, are recommended to completely deactivate Zymit protease activity.
AB - 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 5mM. 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 0.1M. Wash steps with 0.1% SDS were also ineffective in completely removing the remaining Zymit activity. Protocols including wash steps with at least 0.1M NaOH, as for example specified in the original ZENM protocol, are recommended to completely deactivate Zymit protease activity.
UR - https://scripts.iucr.org/cgi-bin/paper?S2053230X14026053
UR - http://www.scopus.com/inward/record.url?scp=84921714835&partnerID=8YFLogxK
U2 - 10.1107/S2053230X14026053
DO - 10.1107/S2053230X14026053
M3 - Article
SN - 1744-3091
VL - 71
SP - 100
EP - 102
JO - Acta Crystallographica Section F:Structural Biology Communications
JF - Acta Crystallographica Section F:Structural Biology Communications
ER -