CSF 14-3-3 Western blot

 

csf

Results

Lanes 1, 2, 3 are positive for CSF 14-3-3

Lanes 4, 5, 6 are weakly positive for CSF 14-3-3

Lanes 7, 8 and 9 are negative for CSF 14-3-3

Lanes 1 and 2 are from two patients with sporadic Creutzfeldt-Jakob disease (CJD)

Lanes 3 and 4 are from patients who have had a stroke

Lanes 5, 6, 7, 8 and 9 contain CSF samples from patients who do not have CJD

In the United Kingdom weakly positive CSF 14-3-3 results are not considered to be supportive of a diagnosis of sporadic CJD.

Method
SDS-PAGE gel:
12% SDS-PAGE running gel with 4% stacking gel
CSF volume:
15μl of CSF sample per well
Electrophoretic conditions:
40mA constant current for 1 hour 20 minutes
Transfer conditions:
0.72mA/cm2 gel for 2 hours
Primary antibody:
Mouse monocional anti-14-3-3b (Santa Cruz Code: sc-1657)1: 1000 dilution incubated overnight
Secondary antibody:
Rabbit anti-mouse lg HRP (DAKO Code:P0260)1: 1000dilution for 1 hour
ECl reagent:
ECL Plus (GE Healthcare Code: RPN2132)
Exposure time:
2 minutes

For more information please contact Alison Green alison.green@ed.ac.uk

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For more information please contact Richard Knight r.knight@ed.ac.uk