This definition appears rarely
Protein denaturation was obtained after an overnight (ON) incubation at RT. Proteins were separated by SDS-PAGE39 and stained with colloidal Coomassie G-25040 or transferred to 0. 45 µm nitrocellulose membrane (Whatman, Springfield, UK)41. For Western blotting, membranes were saturated with Odyssey blocking buffer (LI-COR Biosciences, Lincoln, MA) and incubated with specific antibody according to manufacture instructions: Anti-CD63 (Santa Cruz Biotechnologies, Santa Cruz, CA); anti-ALIX (Thermo Scientific, Waltham, Ma), anti-tumour suppressor gene (TSG101), anti-ß-actin and anti-podocin (Sigma Aldrich; Dorset, UK); anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Abcam, Cambridge, UK); anti dipeptidyl dipeptidase 4 (DDP4 and anti-neprilysin (NEP) (R & D System Inc. , Minneapolis, MN); anti-human IgA, polyclonal Rabbit anti-human alpha-1-antitrypsin, anti-human alpha-2-macroglobulin; anti-ubiquitin, anti-IgG, anti-IgM, polyclonal rabbit anti-human serum albumin (HSA) (Dako, Glostrup, Denmark), mouse anti-HSA (Medix Biochemica, Kauniainen, Finland) anti-non-catalytic region of tyrosine kinase adaptor protein 1 [Nck1] and rabbit anti-Tamm-Horsfall glycoprotein.