20190926 IgG SH-SY5Y neurons in CNTN1/Caspr+ CIDP patients

Samples:

1. 17595 1:100
2. 16047 1:50
3. Regine 1:100
4. Vieira 1:100
5. Piffort 1:100
6. 19186 1:100
7. negative control 1:100

Objective: to test IgG reactivity against SH-SY5Y neuroblastoma derived neurons in in CNTN1/Caspr+ CIDP patients

ICC protocol:

• Fix diferenciated SH-SY5Y cells with 4% PFA 5 min
• Wash 2 times with PBS 1x
• Block with goat serum 5% for 1h at RT
• Dilute serum 1/100 in blocking solution, incubate for 1h
• Wash with PBS 1x
• Commercial anti-neurofascin Ab 1/200 diluted in blocking solution, incubate for 1h
• Wash with PBS 1x
• Secondary antibodies: GAH 594 (1/500), GAC 488 (1/500)
• Wash with PBS 1x (not water)
• Vectashield mounting medium
  

Results: 

1. 17595 1:100 –> neurons’ bodies staining, and mild reactivity against axons
2. 16047 1:50 –> axonal staining
3. Regine 1:100 –> axonal staining
4. Vieira 1:100 –> axonal staining
5. Piffort 1:100 –> axonal staining
6. 19186 1:100 –> –> axonal staining
7. negative control 1:100 –> negative