Author: Pattyn, Els; Verhee, Annick; Uyttendaele, Isabel; Piessevaux, Julie; Timmerman, Evy; Gevaert, Kris; Vandekerckhove, Joël; Peelman, Frank; Tavernier, Jan
Title: HyperISGylation of Old World Monkey ISG15 in Human Cells Document date: 2008_6_18
ID: 1eksm537_35
Snippet: TAP purification experiments were performed in 2fTGH cells stably expressing either Hu or AgmISG15 and HekT cells transiently expressing either Hu or AgmISG15 with an N-terminal TAP-tag. Single colonies of the 2fTGH cells were selected upon co-transfection of the ISG15 constructs with a puromycin resistance marker and selection on 3 mg/mL puromycin. In order to check the expression levels of the TAP-ISG15 proteins compared to the endogenous ISG15.....
Document: TAP purification experiments were performed in 2fTGH cells stably expressing either Hu or AgmISG15 and HekT cells transiently expressing either Hu or AgmISG15 with an N-terminal TAP-tag. Single colonies of the 2fTGH cells were selected upon co-transfection of the ISG15 constructs with a puromycin resistance marker and selection on 3 mg/mL puromycin. In order to check the expression levels of the TAP-ISG15 proteins compared to the endogenous ISG15 levels, the different stable cell-lines were stimulated for 26 h with IFNb (Peprotech) prior to Western blot analysis.
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