Objective: To test reactivity against Purkinje cells in NF155+ CIDP patients. We are going to perform it with two different protocols to decide which one we have to us to test all NF155+ CIDP samples.
Samples: 1.1, 2.1, 3.1, 4.1, 5.1
PROCEDURE:
CONDITION 1: NOVA Lite Monkey Cerebellum/Cerebrum and mouse stomach (504225-Inova)
- Samples diluted in PBS. 30 min at RT
- Wash 3 times with PBS.
- Secondary Ab FITC IgG Conjugate Primate Absorbed (undiluted). 30 min at RT
- Wash 3 times with PBS.
- Mount with Fluoromount mounting medium.
CONDITION 2:
- Incubate with blocking solution (goat serum 5% in PBS) 1h at RT.
- Incubate 50 microl of sample diluted 1/20 in goat serum 5% in PBS 1h at RT.
- Wash 3 times with PBS 1x.
- Secondary Ab GAH monkey absorbed (1/500) 1 hour at RT.
- Wash 3 times with PBS 1x.
- Mount with Fluoromount mounting medium.
RESULTS:
All samples have a positive staining in Purkinje cells.
The two conditions work similar, but condition 2 is better to make the photos. We will use condition 2 to perform the experiment with the other samples.
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