There is conflicting nomenclature used by different investigators to assign pVHL isoforms: pVHL213 is also known as pVHL30 or pVHL25, while pVHL160 is also referred as pVHL19 or pVHL21. In this study, to prevent this confusion, each isoform will be named according to the number of amino acids of the molecule, that is, pVHL213, pVHL160 and pVHL172.
All recombinant proteins were prepared using the E.coli strain BL21(DE3)pLysS transformed with pET21a-hVHL213, hVHL172 or hVHL160 and induced to express the proteins VHL213(His)6, VHL172(His)6 and VHL160(His)6 with IPTG. Proteins were prepared and purified by Talon affinity chromatography following the manufacturer's instructions (Clontech, Mountain View, CA, USA) as described by Martin et al (2013) (link). The purity of the eluted fractions was assessed by 12.5% SDS–PAGE and silver staining.
All recombinant proteins were prepared using the E.coli strain BL21(DE3)pLysS transformed with pET21a-hVHL213, hVHL172 or hVHL160 and induced to express the proteins VHL213(His)6, VHL172(His)6 and VHL160(His)6 with IPTG. Proteins were prepared and purified by Talon affinity chromatography following the manufacturer's instructions (Clontech, Mountain View, CA, USA) as described by Martin et al (2013) (link). The purity of the eluted fractions was assessed by 12.5% SDS–PAGE and silver staining.
