Fig 1.
Mycobacterial outer membrane proteins can be specifically enriched for by extraction with OBG. Immunoblots probed with antisera recognizing the inner membrane proteins FtsH and EccC5, MOM proteins MctB and OmpATb, and cytosolic proteins MMAR_0752 and GroEL2. (A) Equal amounts of pellet (DP) and supernatant (DS) fractions after solubilization of M. marinum cell envelope (CE) with Sarkosyl (S), Triton X-100 (T100), Genapol X-80 (G), and n-octyl-β-d-glucopyranoside (OBG). (B) Immunoblot of fractions corresponding to equal OD units of total lysate (T), soluble fraction (cyt1 27k), and pellet fraction (cell envelopes [CE], 27k) after centrifugation at 27,000 × g, after an additional centrifugation step at 100,000 × g (cyt2 100k and P 100k), and detergent pellet (DP) and supernatant (DS) were obtained after solubilization of CE with 1% OBG. For the CBB-stained SDS-PAGE gel, all membrane fractions (last 4 lanes) were loaded in 3-fold excess compared to total and cytosolic fractions.