HUVECs were lysed on ice using Laemmli buffer, as previously described [5 (link)]. Briefly, 20 µg of total protein for each sample was mixed with a 6x loading dye buffer and loaded onto 10% SDS denaturing poly-acrylamide gels. After transferring proteins to a PVDF membrane (10600021 Euroclone), the membrane was blocked with 5% fat-dried milk (EMR180001 Euroclone) and incubated with primary polyclonal rabbit anti-TMEM230 (1:2500, 21466-1-AP, Proteintech, Rosemont, IL, USA,) and polyclonal goat anti-Lamin A/C (sc 376248 Santa Cruz Biotechnology, Dallas, TX, USA) used as endogenous control at concentration 1:7500. Donkey anti-rabbit (1:20,000, NA934V, Amersham, Cologno Monzese, Mi, Italy) and sheep anti-mouse (1:10,000, NA931V Amersham) were used as secondary antibodies.
Cocola C., Abeni E., Martino V., Piscitelli E., Pelucchi P., Mosca E., Chiodi A., Mohamed T., Palizban M., Porta G., Palizban H., Nano G., Acquati F., Bruno A., Greve B., Gerovska D., Magnaghi V., Mazzaccaro D., Bertalot G., Kehler J., Balbino C., Arauzo-Bravo M.J., Götte M., Zucchi I, & Reinbold R.A. (2024). Transmembrane Protein TMEM230, Regulator of Glial Cell Vascular Mimicry and Endothelial Cell Angiogenesis in High-Grade Heterogeneous Infiltrating Gliomas and Glioblastoma. International Journal of Molecular Sciences, 25(7), 3967.
Corresponding Organization : Institute of Biomedical Technologies
Other organizations :
University of Milan, University Hospital Münster, University of Insubria, IRCCS Policlinico San Donato, MultiMedica, Ospedale Santa Chiara, University of Trento, National Institutes of Health, National Institute of Diabetes and Digestive and Kidney Diseases, University of the Basque Country
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