Objective: To test MS patients for IgG positivity towards FLOT1/FLOT2.
Samples: 101-125 and CTRL –
- Transfection date: 2018.11.08
- Blocking solution: 5% goat serum in PBS
- Patient’s sera diluted 1/100 for IgG with goat serum 5% in PBS.
- Comercial anti-flotillin2 dilluted 1/200 (1h)
- Secondary antibody: GAR488 + GAH 594 IgG (diluted 1/1000)
- Vectashield mounting medium
RESULTS: Many of the samples were positive, but just the cells that seems to be death (stained with FLOT2). We think that there is a problem with the transfection. We are going to try a new transfeccion but not leaving the cells for 48h as the original paper; we are going to leave the cells just for 36 hours.
References
- Hahn S1, Trendelenburg G2, Scharf M1, Denno Y1, Brakopp S1, Teegen B1,3, Probst C1, Wandinger KP4, Buttmann M5,6, Haarmann A5, Szabados F7, Vom Dahl M8, Kümpfel T9, Eichhorn P10, Gold H11, Paul F12,13, Jarius S14, Melzer N15, Stöcker W1,3, Komorowski L16. Identification of the flotillin-1/2 heterocomplex as a target of autoantibodies in bona fide multiple sclerosis. J Neuroinflammation. 2017 Jun 23;14(1):123. PMID: 28645295.
