Bitan:Extraction of mice brains for α-syn ELISA

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Brain Extraction Protocol

Brain Extraction Protocol

-       Make lysis buffer; keep on ice

o      1 tablet Com antiphos into 1 mL lyis buffer = 25X stock 40ul/1mL

o      10mL lysis buffer + 1 phostop tablet + 40uL Com

o      10mL lysis + 1% triton

-       Get brains, keep on ice, weigh scale in B117B

-       Put brains in 5mL glass tube

-       1mL lysis + phsph + Com into tube

-       Get beakers of ddH2O and 100% EtOH

o      Use pellet/pestle motor to crush

o      Clean tip in water at EtOH and wipe between brains

-       Sonicate on low (~1 or 2) till looks homogenous

o      keep on ice while sonicating

o      sonicate in bursts otherwise it heats solution and can denature protein

o      transfer to 1.5mL tube (might need dropper for chunks)

§       might want to wait 10-15 min before spinning

-       Spin

-       Collect

-       Freeze all samples

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