20150306 Dot_blot human_nerve_lysate

Objective

We’ll try to see positive marks for FXM that it has antibodies against a more abundant protein of the human nerve, PMP2.

We also want to test purified IgG and other positive patients as LAC and JML which are positive for neurofascin (strong positives of a non-abundant protein; different from FXM).

Material

see the last dot_blot

lysate of human nerve with crosslink buffer
lysate of human nerve with crosslink buffer and washed with protein A/G

Methods

see the last dot_blot

Changes:

Put a drop of each lysate in the same membrane
Sera incubation 1/100 in a wet chamber
Purified IgG also 1/100 in a wet chamber

Distribution:

JML_IgG
FXM_IgG
JML
LAC
Ctrl
FXM

Results

Figure 1.Dot blot with human lysate with crosslink buffer 1. JML_IgG 2. FXM_IgG 3. JML 4. LAC 5. Ctrl 6.FXM — Figure 1.Dot blot with human lysate with crosslink buffer
1. JML_IgG
2. FXM_IgG
3. JML
4. LAC
5. Ctrl
6.FXM

Figure 2. Dot blot with human lysate with crosslink buffer after protein A/G 1. JML_IgG 2. FXM_IgG 3. JML 4. LAC 5. Ctrl 6. FXM — Figure 2. Dot blot with human lysate with crosslink buffer after protein A/G
1. JML_IgG
2. FXM_IgG
3. JML
4. LAC
5. Ctrl
6. FXM

Figure 3.Dot blot with human lysate with lysis buffer 1. JML_IgG 2. FXM_IgG 3. JML 4. LAC 5. Ctrl 6. FXM — Figure 3.Dot blot with human lysate with lysis buffer
1. JML_IgG
2. FXM_IgG
3. JML
4. LAC
5. Ctrl
6. FXM

We cannot see positive marks in any dot.

Conclusions

We’ll change the experiment. Our new approach for the screening is to label purified IgG from patients with fluorescence and then do an IHC with human nerve.

20150306 Dot_blot human_nerve_lysate

Published by Roser Alba

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