20190926: Monkey’s peripheral nerve IHC in CIDP patients

Objective: To investigate Monkey’s Peripheral nerve IHC staining paterns of CIDP patients with CNTN1/Caspr1 Ab, a colocalization with Caspr1 commercial mAb

Samples:

1.  negative control 1:10
2. MRVM 1:20
3. Vieira 1:20
4. Piffort 1:20
5. Regine 1:20

IHQ protolocol:

• We used commercial slides from Inova (peripheral nerve)
• Incubate with blocking solution (goat serum 5% in PBS) 1h
• Incubate 50 microl of sample (non-absorbed) 1h
• Wash 3 times with PBS 1x.
• Incubate with Caspr1 commercial mAb (1:20)
• Secondary Ab GAH AF488 monkey absorbed  (1/400), GAR 594 for 2. 1 hour at RT.
• Wash 3 times with PBS 1x.
• Mount with Fluoromount mounting medium.

Results:

1.  negative control 1:10 –> negative
2. MRVM 1:20 –> a lot of background
3. Vieira 1:20 –> amielinic fibers and paranodes?
4. Piffort 1:20 –> paranodes, colocalization with Caspr1
5. Regine 1:20 –> paranodes, colocalization with Caspr1