Α-syn



       &#945;-synuclein



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  &#945;-synuclein</o:p> 

<p class=MsoNormal>Protein Concentration: 20&#956;M

<p class=MsoNormal>Protocol:

<ul style='margin-top:0in' type=disc> <li class=MsoNormal style='mso-list:l0 level1 lfo3;tab-stops:list .5in'>Incubate &#945;-syn in the differential media at a concentration of 200 &#956;M for 24 h prior to adding to the cells. The protein is very soluble at this concentration. </li> <li class=MsoNormal style='mso-list:l0 level1 lfo3;tab-stops:list .5in'>After 24h, add 10 &#956;l of this solution to the cells. </li> <li class=MsoNormal style='mso-list:l0 level1 lfo3;tab-stops:list .5in'>Incubate for 24h and do the MTT assay (48h for LDH assay)</li> <li class=MsoNormal style='mso-list:l0 level1 lfo3;tab-stops:list .5in'>If you want to do dose dependence with a drug, prepare the dilutions of the drug in the media in 70 &#956;l volume (for 6 replicates). Add 70 &#956;l of    200 &#956;M &#945;-syn to the drug solution. Take out 10 &#956;l of media from the plated cells and add 20 &#956;l of the peptide+ drug to the cells (6 replicates). Do MTT/LDH as required.</li>

<li class=MsoNormal style='mso-list:l0 level1 lfo3;tab-stops:list .5in'>The viability of 20&#956;M &#945;-syn is ~60-70% by MTT</li> </ul>

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