Calcitonin



       &#946;2-micoglobulin



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  Calcitonin 

Protein Concentration: 15&#956;M

<p class=MsoNormal>Cells: PC12 cells

<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 calcitonin in MiliQ water at a concentration of 1.5 mM 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'>Dilute this solution to 150 &#956;M in differential media after 24h and 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.</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    150 &#956;M calcitonin 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 15&#956;M calcitonin is ~70-80% by MTT assay</li> </ul>

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