Samples:
- GBS IGOS 51-99
- Negative control
- Positive control (GBS IGOS 5)
Protocol:
- Cells seeded on 23.01.20, differentiated on 24.01.20 and 27.01.20
- Fix for 15 minutes with PFA 4 %
- Wash 3 times with PBS1x
- Block for 1 hour with Goat Serum 5 %
- Patient’s sera
- For IgM: diluted 1/40 in Goat serum 5 % (1 h)
- Wash 3 times with PBS1x
- Commercial antibody: anti-NF (AF3235) diluted 1/200 in Goat serum 5% (1h)
- Wash 3 times with PBS1x
- Secondary antibodies: GAC594 IgG + GAH488 IgM diluted 1/500 in Goat serum 5 % (1 h)
- Wash 3 – 6 times with PBS1x (NO H2Od)
- Vectashield mounting medium
RESULTS:
- 53 ++
- 58 +
- 62, 63 repetir
- 65 +
- 67 +
- 71 ++
- 75 +
- 77 +
- 78 +++
- 79 +++
- 85 +
- 87 ++ / +++
- 94 +
- 95 +++
- 96 +
- 98 +
- 5 +++
