SurA | SurA
AS10 798 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Yersinia pseudotuberculosis
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Recombinant SurA over-expressed as a N-terminal fusion with His(x6)-tag, which was removed prior to immunization. UniProt protein accession number B1JKY1; NCBI annotated locus tag YPK_3571
Approximately 10 µg of total protein from Y. pseudotuberculosis extracted with were separated on 12 % SDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in milk solution (10 % milk powder in TBST) for 1 h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 5 000 overnight at 4 °C with agitation. The antibody solution was decanted and the blot was washed twiced for 15 min in TBST at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from GE Healthcare) diluted to 1: 20 000 in for 1h at room temperature with agitation. The blots were washed as above and developed for 35 min with homemade ECL detection reagent. Exposure time was 60 seconds.
SurA (stationary phase survival) is a periplasmic chaperone and folding catalyst with peptidyl-prolyl cis-trans isomerase activity. It is involved in the maintenance of Gram-negative bacterial envelope integrity and the transport/assembly of a subset of OMPs.
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