Calcitonin

From OpenWetWare
Jump to navigationJump to search

<html xmlns:v="urn:schemas-microsoft-com:vml" xmlns:o="urn:schemas-microsoft-com:office:office" xmlns:w="urn:schemas-microsoft-com:office:word" xmlns="http://www.w3.org/TR/REC-html40">

<head> <meta name=Title content=&#946;2-micoglobulin> <meta name=Keywords content=""> <meta http-equiv=Content-Type content="text/html; charset=macintosh"> <meta name=ProgId content=Word.Document> <meta name=Generator content="Microsoft Word 11"> <meta name=Originator content="Microsoft Word 11"> <link rel=File-List href="cal_files/filelist.xml"> <title>&#946;2-micoglobulin</title> <!--[if gte mso 9]><xml>

<o:DocumentProperties>
 <o:Author>Sharmistha Sinha</o:Author>
 <o:Template>Normal</o:Template>
 <o:LastAuthor>Sharmistha Sinha</o:LastAuthor>
 <o:Revision>2</o:Revision>
 <o:Created>2009-12-12T00:42:00Z</o:Created>
 <o:LastSaved>2009-12-12T00:42:00Z</o:LastSaved>
 <o:Pages>1</o:Pages>
 <o:Words>107</o:Words>
 <o:Characters>615</o:Characters>
 <o:Company>Neuro</o:Company>
 <o:Lines>5</o:Lines>
 <o:Paragraphs>1</o:Paragraphs>
 <o:CharactersWithSpaces>755</o:CharactersWithSpaces>
 <o:Version>11.1282</o:Version>
</o:DocumentProperties>
<o:OfficeDocumentSettings>
 <o:AllowPNG/>
</o:OfficeDocumentSettings>

</xml><![endif]--><!--[if gte mso 9]><xml>

<w:WordDocument>
 <w:DisplayHorizontalDrawingGridEvery>0</w:DisplayHorizontalDrawingGridEvery>
 <w:DisplayVerticalDrawingGridEvery>0</w:DisplayVerticalDrawingGridEvery>
 <w:UseMarginsForDrawingGridOrigin/>
</w:WordDocument>

</xml><![endif]--> <style> <!--

/* Font Definitions */

@font-face {font-family:"Times New Roman"; panose-1:0 2 2 6 3 5 4 5 2 3; mso-font-charset:0; mso-generic-font-family:auto; mso-font-pitch:variable; mso-font-signature:50331648 0 0 0 1 0;}

/* Style Definitions */

p.MsoNormal, li.MsoNormal, div.MsoNormal {mso-style-parent:""; margin:0in; margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:12.0pt; font-family:"Times New Roman";} table.MsoNormalTable {mso-style-parent:""; font-size:10.0pt; font-family:"Times New Roman";} @page Section1 {size:8.5in 11.0in; margin:1.0in 1.25in 1.0in 1.25in; mso-header-margin:.5in; mso-footer-margin:.5in; mso-paper-source:0;} div.Section1 {page:Section1;}

/* List Definitions */

@list l0 {mso-list-id:827212806; mso-list-type:hybrid; mso-list-template-ids:-1404653784 67698689 67698691 67698693 67698689 67698691 67698693 67698689 67698691 67698693;} @list l0:level1 {mso-level-number-format:bullet; mso-level-text:\F0B7; mso-level-tab-stop:.5in; mso-level-number-position:left; text-indent:-.25in; font-family:Symbol;} ol {margin-bottom:0in;} ul {margin-bottom:0in;} --> </style> <!--[if gte mso 9]><xml>

<o:shapedefaults v:ext="edit" spidmax="1027"/>

</xml><![endif]--><!--[if gte mso 9]><xml>

<o:shapelayout v:ext="edit">
 <o:idmap v:ext="edit" data="1"/>
</o:shapelayout></xml><![endif]-->

</head>

<body bgcolor=white lang=EN-US style='tab-interval:.5in'>

<div class=Section1>

<p class=MsoNormal><span style='font-size:14.0pt'><b><![if !supportEmptyParas]>&nbsp;<![endif]><o:p></o:p></b></span></p>

<p class=MsoNormal><span style='font-size:14.0pt'><b><u>Calcitonin<o:p></o:p></u></b></span></p>

<p class=MsoNormal>Protein Concentration: 15&#956;M</p>

<p class=MsoNormal>Cells: PC12 cells</p>

<p class=MsoNormal>Protocol: </p>

<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>

<p class=MsoNormal><![if !supportEmptyParas]>&nbsp;<![endif]><o:p></o:p></p>

</div>

</body>

</html>