Brain extracts from Xenopus tadpoles



Prepare extraction buffer + Protease Inhibitors and keep tube on ice, prepare 5l buffer/brain


Dissect brains and collect them in microtube with extraction buffer on ice, try to transfer as little liquid as possible with the brains


Use a 200l pipet tip which got squeezed on the tip to create a very thin opening

Homogenize the brains by pipetting up and down until the brains are smashed

If it takes a long time to homogenize, put the sample on ice in between


Spin the homogenate 5min at 13000 rpm at 4C


Collect the Supernatant SN and transfer to a new tube, add 2x Sample Buffer SB + DTT


Incubate 5min at 100C


Freeze samples at -20C



Extraction Buffer:


10mM HEPES/NaOH pH 7.4

150mM NaCl

2mM EDTA pH 8.0

1% NP40


store at 4C



Protease Inhibitors:

Calbiochem Protease Inhibitor Set V, EDTA-free

100x solution, store at -20

add Protease Inhibitors freshly before each extraction