Objective: To investigate Monkey’s Cerebellum and Cerebrum IH staining paterns of CIDP patients with CNTN1/Caspr1 Ab
Samples: negative control, 17595, 15198 (CNTN1+)
Numbers:
- 1 – negative control Pre-absorbed
- 2 – 17595 Pre-absorbed
- 3 – 17595
- 4 – 15198 Pre-absorbed
- 5 – 15198
We used commercial slides from Euroimmune (brain and peripheral nerve)
Serum absorption for immunohistochemistry
- Defreeze an eppendorf of the box “Nucleos para IHQ Monkey” stored at -20ºC.
- Dilute serum in PBS 1x at 1:10. Total volumen 50 microl (5:50). Final dilution: 1:20
- Mix diluted serum with 50microl of diluted HEK cells.
- Agitate in an orbital agitator at RT for two hours (alternative: over night at 4ºC).
IHQ protolocol:
- Incubate with blocking solution (goat serum 5% in PBS) 1h
- Incubate 100 microl of sample (pre-absorbed or non-absorbed)
- Wash 3 times with PBS 1x.
- Secondary Ab GAH monkey absorbed (1/200). 1 hour at RT.
- Wash 3 times with PBS 1x.
- Mount with Fluoromount mounting medium.
RESULTS:
No differences in patterns between pre-absorbed and non-absorbed samples were observed.
Nerve:
– 17595: nonspecific staining of axons and interstices that prevents the paranodes from being seen
– 15198: staining of paranodes and patched areas which could correspond to amielinic fibres
Brain:
– 17595: staining around purkinje cells.
– 15198: neuropilo pattern.
