Fig. 2.

Large-scale purification of membrane NEU1 and identification of already know interaction partners. a Left panel: 4.8 × 10⁸ macrophages differentiated from THP-1 cells were submitted to the purification protocol and proteins recovered at the end of the purification were separated by SDS-PAGE and stained by Coomassie Blue (lane 2). Lane 1, protein lysate (starting material); lane 3, proteins recovered after elution with Laemmli buffer of protein G Sepharose beads coated with anti-NEU1 antibodies only. Asterisks, non-specific protein bands recovered in both lanes 2 and 3. Right panel: as illustrated for lane 2, lanes were excised into 23 bands and in-gel digested by trypsin for analysis by mass spectrometry. b Identification by LC–MS/MS of already know interaction partners of membrane NEU1. Trypsin-digested protein bands were analyzed by nano-LC–ESI MS/M, and proteins identified with Mascot software in Swiss-Prot and trEMBL databases