20191107 IgG SH-SY5Y neurons in CNTN1/Caspr+ CIDP patients

Samples:

1. negative control
2. MRVM (SAMPLE 78 BOX CIDP)
3. 19209
4. 17595
5. 19204
6. 19212
7. Vieira
8. 19216
9. CNTN1+ 15198

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 15 min
• Wash 2 times with PBS 1x
• Block with rabbit serum 1/40 for 1h at RT
• Dilute serum 1/50 in blocking solution, incubate for 1h
• Wash with PBS 1x
• Commercial anti-CNTN1 Ab 1/00 diluted in blocking solution, incubate for 1h
• Wash with PBS 1x
• Secondary antibodies: RAH 594 (1/500), RAG 488 (1/500)
• Wash with PBS 1x (not water)
• Vectashield mounting medium
  

Results:

1. negative control –> NEGATIVE (somas mild +, axons -)
2. MRVM (SAMPLE 78 BOX CIDP) –> MILD POSITIVE (somas + axons)
3. 19209 –> MILD POSITIVE (somas + axons)
4. 17595 –> MILD POSITIVE (somas + axons)
5. 19204 –> MILD POSITIVE (somas + axons)
6. 19212 –> MILD POSITIVE (somas + axons)
7. Vieira –> MILD POSITIVE (somas + axons)
8. 19216 –> MILD POSITIVE (somas + axons)
9. CNTN1+ 15198 –> POSITIVE ++ (somas + axons)