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Western Blotting

VN Valerio Napolioni
MP Marilena Pariano
MB Monica Borghi
VO Vasilis Oikonomou
CG Claudia Galosi
AL Antonella De Luca
CS Claudia Stincardini
CV Carmine Vacca
GR Giorgia Renga
VL Vincenzina Lucidi
CC Carla Colombo
EF Ersilia Fiscarelli
CL Cornelia Lass-Flörl
AC Alessandra Carotti
LD Lucia D'Amico
FM Fabio Majo
MR Maria Chiara Russo
HE Helmut Ellemunter
AS Angelica Spolzino
PM Paolo Mosci
SB Stefano Brancorsini
FA Franco Aversa
AV Andrea Velardi
LR Luigina Romani
CC Claudio Costantini
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Cells were lysed in 2x Laemmli buffer (Sigma-Aldrich). Blots of cell lysates were incubated with an antibody against IDO1 (clone 10.1, Millipore) followed by IgG–HRP-conjugated secondary antibody (Sigma–Aldrich) after separation in 10 or 12% Tris/glycine SDS gel and transferred to a nitrocellulose membrane. A cell lysate of IFNγ-stimulated HeLa cells was used as positive control in selected experiments for correct assignment of IDO1 band. Normalization was performed probing the membrane with mouse-anti-β-tubulin antibody (Sigma–Aldrich, clone T9026). Chemiluminescence detection was performed with LiteAblotPlus chemiluminescence substrate (Euroclone S.p.A), using the ChemiDocTM XRS+Imaging system (Bio-Rad), and quantification was obtained by densitometry image analysis using Image Lab 6.0 software (Bio-Rad).

Copyright and License information:  ©2019 Napolioni, Pariano, Borghi, Oikonomou, Galosi, De Luca, Stincardini, Vacca, Renga, Lucidi, Colombo, Fiscarelli, Lass-Flörl, Carotti, D'Amico, Majo, Russo, Ellemunter, Spolzino, Mosci, Brancorsini, Aversa, Velardi, Romani and Costantini.
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