Bitan:Extraction of mice brains for α-syn ELISA
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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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