Objectives
To test if patients’ sera react against diferent protein present in the node of Ranvier.
Materials
- Transfected cells grown in coverslips
- PBS
- H2O
- Goat5%
- GAH488
- GAR488
- GAM488
- Patients sera
Methods
| CNTN1-H | |||
| mouse-anti-CNTN 1/500 | GAM488 | ||
| rabbit-anti-CNTN 1/120 | GAR488 | ||
| HVH1 | GAH488 | ||
| 104 | GAH488 | ||
| CD9 | |||
| CLG | GAH488 | ||
| COC | GAH488 | ||
| AR | GAH488 | ||
| ACB | GAH488 | ||
| JPL | GAH488 | ||
| MPG | GAH488 | ||
| ctrl7 | GAH488 | ||
| Xavi | GAH488 | ||
| 2 Esther | GAH488 | ||
| 2 | GAH488 | ||
| 2- | GAH488 | ||
| Ana | GAH488 | ||
| NF186 | |||
| CLG | rat-anti-HA 1/2000 | GAH-594 + GARat-488 | |
| TMF | rat-anti-HA 1/2000 | GAH-594 + GARat-488 | |
| HVH1 | rat-anti-HA 1/2000 | GAH-594 + GARat-488 | |
| ATR | rat-anti-HA 1/2000 | GAH-594 + GARat-488 | |
| MRVM | rat-anti-HA 1/2000 | GAH-594 + GARat-488 | |
| RVT | rat-anti-HA 1/2000 | GAH-594 + GARat-488 | |
| RMM | rat-anti-HA 1/2000 | GAH-594 + GARat-488 | |
| JGV | rat-anti-HA 1/2000 | GAH-594 + GARat-488 | |
| MOE | rat-anti-HA 1/2000 | GAH-594 + GARat-488 | |
| LDC | rat-anti-HA 1/2000 | GAH-594 + GARat-488 | |
| MASP | rat-anti-HA 1/2000 | GAH-594 + GARat-488 | |
| JHH | rat-anti-HA 1/2000 | GAH-594 + GARat-488 |
- Incubation time: 1 hour
- Wash 3 times with PBS between each incubation
Results & Conclusions
CNTN1-Human (Genecopoeia)
There are very positive marks for HVH1, therefore the cells are transfected. But there aren’t any positive marks for commercial antibodies (mouse + rabbit), at least anything like HVH1. Patients 104 (from CNTN-NFASC data base) is negative.
CD9
This time cells were not permeabilized. All 6 patients and 6 controls are “clearly” negative.
NF186
There are positive marks for Ha tag, therefore the cells are transfected. None of the 12 patients are positive for NF186.
