TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<div title="Page 1">
<div>
<div>
<p>Protocol for adult (8-10 week) mouse kidney dissociation.</p>
</div>
</div>
</div>

[BEFORE_START]
<div title="Page 1">
<p>Prepare <em>Bacillus Licheniformis</em> enzyme mix just prior to starting dissociation:</p>
<p> </p>
<table style="height: 132px; width: 552px;">
<tbody>
<tr>
<td style="width: 145px; text-align: left;"><strong>Volume (µl)</strong></td>
<td style="width: 206px; text-align: left;"><strong>Reagent</strong></td>
<td style="width: 179px; text-align: left;"><strong> Final concentration </strong></td>
</tr>
<tr>
<td style="width: 145px;"> 894</td>
<td style="width: 206px;"> DPBS</td>
<td style="width: 179px;"> 1X</td>
</tr>
<tr>
<td style="width: 145px;"> 1</td>
<td style="width: 206px;"> 0.5 M EDTA</td>
<td style="width: 179px;"> 0.5 mM</td>
</tr>
<tr>
<td style="width: 145px;"> 5</td>
<td style="width: 206px;"> DNAse 1 (250 U/10 μL)</td>
<td style="width: 179px;"> 125 U / mL</td>
</tr>
<tr>
<td style="width: 145px;"> 100</td>
<td style="width: 206px;"> <em>B. Lich</em> (100 mg/mL)</td>
<td style="width: 179px;"> 10 mg/mL</td>
</tr>
</tbody>
</table>
<div title="Page 1">
<div>
<div>
<p><strong>+25 mg tissue / 1 mL enzyme mix</strong></p>
</div>
</div>
</div>
</div>

[GUIDELINES]
<p><strong>Storage Conditions of Reagents </strong></p>
<p> </p>
<table style="height: 156px; width: 554px;">
<tbody>
<tr>
<td style="width: 216px; text-align: left;">
<div title="Page 1">
<div>
<div>
<div>
<p><strong>Reagent</strong></p>
</div>
</div>
</div>
</div>
</td>
<td style="width: 326px; text-align: left;">
<div title="Page 1">
<p><strong>Storage Condition </strong></p>
</div>
</td>
</tr>
<tr>
<td style="width: 216px;">
<div title="Page 1">
<div>
<div>
<div>
<p>DPBS (no Ca, no Mg)</p>
</div>
</div>
</div>
</div>
</td>
<td style="width: 326px;">
<div title="Page 1">
<p>4°C</p>
</div>
</td>
</tr>
<tr>
<td style="width: 216px;">
<div title="Page 1">
<div>
<div>
<div>
<p>0.5 M EDTA</p>
</div>
</div>
</div>
</div>
</td>
<td style="width: 326px;">
<div title="Page 1">
<div title="Page 1">
<div>
<div>
<div>
<p>room temp.</p>
</div>
</div>
</div>
</div>
</div>
</td>
</tr>
<tr>
<td style="width: 216px;">
<div title="Page 1">
<div>
<div>
<div>
<p>RBC Lysis Buffer</p>
</div>
</div>
</div>
</div>
</td>
<td style="width: 326px;">4°C</td>
</tr>
<tr>
<td style="width: 216px;">
<div title="Page 1">
<div>
<div>
<div>
<p>Protease from <em>Bacillus Licheniformis</em></p>
</div>
</div>
</div>
</div>
</td>
<td style="width: 326px;">
<div title="Page 1">
<div>
<div>
<div>
<p>Store 100 μL aliquots (100 mg/mL) in DPBS at -80°C</p>
</div>
</div>
</div>
</div>
</td>
</tr>
<tr>
<td style="width: 216px;">
<div title="Page 1">
<div>
<div>
<div>
<p>DNAse</p>
</div>
</div>
</div>
</div>
</td>
<td style="width: 326px;"> Store 10 μL aliquots (250 U/10 μL) in DPBS at -80°C</td>
</tr>
</tbody>
</table>
<p> </p>
<p> </p>
<div title="Page 1">
<div>
<div>
<p><strong>Required Equipment</strong></p>
<p> </p>
<table style="height: 46px; width: 553px;">
<tbody>
<tr>
<td style="width: 167px; text-align: left;">
<div title="Page 1">
<div>
<div>
<div>
<p><strong>Equipment</strong></p>
</div>
</div>
</div>
</div>
</td>
<td style="width: 141px; text-align: left;">
<div title="Page 1">
<div>
<div>
<div>
<p><strong>Supplier</strong></p>
</div>
</div>
</div>
</div>
</td>
<td style="width: 223px; text-align: left;">
<div title="Page 1">
<div>
<div>
<div>
<p><strong>Catalog no.</strong></p>
</div>
</div>
</div>
</div>
</td>
</tr>
<tr>
<td style="width: 167px;">
<div title="Page 1">
<div>
<div>
<div>
<p>Thermomixer C or R</p>
</div>
</div>
</div>
</div>
</td>
<td style="width: 141px;">
<div title="Page 1">
<div>
<div>
<div>
<p>Eppendorf</p>
</div>
</div>
</div>
</div>
</td>
<td style="width: 223px;">
<div title="Page 1">
<div>
<div>
<div>
<p><a href="https://online-shop.eppendorf.com/OC-en/eshopproduct/view/5382000015?_ga=2.260930253.1154207520.1519596651-1488113111.1519596651" target="_blank">5382000015</a> / <a href="https://www.sigmaaldrich.com/catalog/product/sigma/z605271?lang=en&amp;region=US" target="_blank">Z605271</a></p>
</div>
</div>
</div>
</div>
</td>
</tr>
</tbody>
</table>
<p> </p>
<p> </p>
<p><strong>The protocol workflow is as follows: </strong></p>
<p> </p>
<div title="Page 1">
<div>
<div>
<p>A. Isolate Kidney</p>
<div title="Page 1">
<div>
<div>
<p>B. First layer</p>
<div title="Page 2">
<div>
<div>
<p>C. Second layer</p>
<div title="Page 2">
<div>
<div>
<p>D. Third layer</p>
<div title="Page 2">
<div>
<div>
<p>F. Preparing cells for Chromium</p>
</div>
</div>
</div>
</div>
</div>
</div>
</div>
</div>
</div>
</div>
</div>
</div>
</div>
</div>
</div>
</div>
</div>
</div>

[STEPS]
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?.