Rnasealert kit

Manufactured by Thermo Fisher Scientific

The RNaseAlert kit is a fluorescence-based detection system designed to monitor the presence of RNase, a ubiquitous enzyme that can degrade RNA samples. The kit provides a simple and effective way to detect RNase contamination in laboratory settings.

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Lab products found in correlation

Q5 site directed mutagenesis kit, by New England Biolabs (1 mentions)

Close spelling variants of this product

RNaseAlert Lab Test Kit Ambion RNaseAlert Lab Test kit

2 protocols using rnasealert kit

Fab Protein Expression and Purification

Cited 1 time

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Output clones were sequenced and reformatted with the introduction of stop codon on phagemids using Q5 site-directed mutagenesis kit (NEB). The desired Fab proteins were then expressed as soluble proteins according to published protocols (14 (link),16 (link)). Collected cell pellets were lysed, and Fab proteins were purified using the AKTAxpress fast protein liquid chromatography (FPLC) purification system (Amersham) as described. Purified protein was dialyzed into 1 × PBS (pH 7.4), concentrated and analyzed by 12% sodium dodecyl sulphate-polyacrylamide gel electrophoresis using Coomassie Blue R-250 staining for visualization. Aliquots of Fab samples were tested for RNase activity using the RNaseAlert kit (Ambion). The aliquots of Fab samples were flash frozen in liquid nitrogen and stored at −80°C until further use. Additional methods and requests for material including Fab BL3–6 and Fab BL3–6S97N (if available), Fab plasmids and the detailed protocols for expression and purification of Fabs will be provided upon request.
Additional details on material and methods including the nucleic acid sequences are available in Supplementary Data.

Koirala D., Shelke S.A., Dupont M., Ruiz S., DasGupta S., Bailey L.J., Benner S.A, & Piccirilli J.A. (2018). Affinity maturation of a portable Fab–RNA module for chaperone-assisted RNA crystallography. Nucleic Acids Research, 46(5), 2624-2635.

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Recombinant Fab Protein Purification

Cited 2 times

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Output clones were sequenced, and reformatted with introduction of stop codon on phagemids as described^{29 (link)}. The desired Fab proteins were then expressed as soluble proteins according to published protocols^{66 (link)}. Collected cell pellets were lysed, and Fab proteins were purified using the AKTAxpress FPLC purification system (Amersham) as described^{30 (link)}. Purified protein was dialyzed into 1x PBS pH 7.4, concentrated and analyzed by 12% SDS-PAGE using Coomassie Blue R-250 staining for visualization. Aliquots of Fab samples were tested for RNase activity using the RNase-alert kit (Ambion).

Shao Y., Huang H., Qin D., Li N.S., Koide A., Staley J.P., Koide S., Kossiakoff A.A, & Piccirilli J.A. (2016). Specific Recognition of a Single-stranded RNA Sequence by a Synthetic Antibody Fragment. Journal of molecular biology, 428(20), 4100-4114.

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