Objective: To investigate Monkey’s Cerebellum and Cerebrum IH staining paterns of patients’ serum with immune-mediated neuropathies.
Samples: 19-061, NF155 (De Haro), 16-047, MRV, Negative control.
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).
- 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 50microl of sample (absorbed or diluted) 30 min at RT.
- Wash 3 times with PBS 1x.
- Secondary Ab FITC IgG Conjugate Primate Absorbed (1/1000). 30 min at RT.
- Wash 3 times with PBS 1x.
- Mount with Fluoromount mounting medium.
RESULTS: Sample 16-041 (positive for NF155 in ICC but not in IH) don’t have the same pattern staining than a NF155+ (staining of the pukinje cells); its staining is very similar to negative control. Samples 16-047 and MRV have a neuropil pattern (similar to CNTN1 patients).
