Sedimentation Densities
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Materials
Table2 Density and refractive indexes of sucrose
Table1 Densities of cell structures in sucrose
Sucrose density fractions from Exercise “Sucrose Fractionation of Castor Bean”
Procedure
Using the densities for sucrose given in Table 2 , compute the density of each organelle found within the fractions collected by differential centrifugation.
Compare these values with those listed in Table 1
Notes
It is possible to approximate the densities of the particles separated above, based on the linear gradient separation data. It would be better to perform an additional centrifugation using CsCl as the viscous media, but for our purposes, the density of the sucrose will be sufficient.
<center><strong>Table 2 Density and refractive indexes of sucrose</strong></center><center></center><center></center><center></center><center> <table> <tbody> <tr> <th>Density (g/cm<sup>3</sup>)</th> <th>Refractive index</th> <th>% by weight (w/v)</th> <th>Molarity</th> </tr> <tr> <td>0.9982</td> <td>1.3330</td> <td>0</td> <td><br /></td> </tr> <tr> <td>1.0021</td> <td>1.3344</td> <td>1</td> <td>0.029</td> </tr> <tr> <td>1.0060</td> <td>1.3359</td> <td>2</td> <td>0.059</td> </tr> <tr> <td>1.0099</td> <td>1.3374</td> <td>3</td> <td>0.089</td> </tr> <tr> <td>1.0139</td> <td>1.3388</td> <td>4</td> <td>0.119</td> </tr> <tr> <td>1.0179</td> <td>1.3403</td> <td>5</td> <td>0.149</td> </tr> <tr> <td>1.0219</td> <td>1.3418</td> <td>6</td> <td>0.179</td> </tr> <tr> <td>1.0259</td> <td>1.3433</td> <td>7</td> <td>0.210</td> </tr> <tr> <td>1.0299</td> <td>1.3488</td> <td>8</td> <td>0.211</td> </tr> <tr> <td>1.0340</td> <td>1.3464</td> <td>9</td> <td>0.272</td> </tr> <tr> <td>1.0381</td> <td>1.3479</td> <td>10</td> <td>0.303</td> </tr> <tr> <td>1.0423</td> <td>1.3494</td> <td>11</td> <td>0.335</td> </tr> <tr> <td>1.0465</td> <td>1.3510</td> <td>12</td> <td>0.367</td> </tr> <tr> <td>1.0507</td> <td>1.3526</td> <td>13</td> <td>0.399</td> </tr> <tr> <td>1.0549</td> <td>1.3541</td> <td>14</td> <td>0.431</td> </tr> <tr> <td>1.0592</td> <td>1.3557</td> <td>15</td> <td>0.464</td> </tr> <tr> <td>1.0635</td> <td>1.3573</td> <td>16</td> <td>0.497</td> </tr> <tr> <td>1.0678</td> <td>1.3590</td> <td>17</td> <td>0.530</td> </tr> <tr> <td>1.0721</td> <td>1.3606</td> <td>18</td> <td>0.564</td> </tr> <tr> <td>1.0765</td> <td>1.3622</td> <td>19</td> <td>0.597</td> </tr> <tr> <td>1.0810</td> <td>1.3639</td> <td>20</td> <td>0.632</td> </tr> <tr> <td>1.0854</td> <td>1.3655</td> <td>21</td> <td>0.666</td> </tr> <tr> <td>1.0899</td> <td>1.3672</td> <td>22</td> <td>0.701</td> </tr> <tr> <td>1.0944</td> <td>1.3689</td> <td>23</td> <td>0.735</td> </tr> <tr> <td>1.0990</td> <td>1.3706</td> <td>24</td> <td>0.771</td> </tr> <tr> <td>1.1036</td> <td>1.3723</td> <td>25</td> <td>0.806</td> </tr> <tr> <td>1.1082</td> <td>1.3740</td> <td>26</td> <td>0.842</td> </tr> <tr> <td>1.1128</td> <td>1.3758</td> <td>27</td> <td>0.878</td> </tr> <tr> <td>1.1175</td> <td>1.3775</td> <td>28</td> <td>0.914</td> </tr> <tr> <td>1.1222</td> <td>1.3793</td> <td>29</td> <td>0.951</td> </tr> <tr> <td>1.1270</td> <td>1.3811</td> <td>30</td> <td>0.988</td> </tr> <tr> <td>1.1318</td> <td>1.3829</td> <td>31</td> <td>1.025</td> </tr> <tr> <td>1.1366</td> <td>1.3847</td> <td>32</td> <td>1.063</td> </tr> <tr> <td>1.1415</td> <td>1.3865</td> <td>33</td> <td>1.100</td> </tr> <tr> <td>1.1463</td> <td>1.3883</td> <td>34</td> <td>1.138</td> </tr> <tr> <td>1.1513</td> <td>1.3902</td> <td>35</td> <td>1.177</td> </tr> <tr> <td>1.1562</td> <td>1.3920</td> <td>36</td> <td>1.216</td> </tr> <tr> <td>1.1612</td> <td>1.3939</td> <td>37</td> <td>1.255</td> </tr> <tr> <td>1.1663</td> <td>1.3958</td> <td>38</td> <td>1.295</td> </tr> <tr> <td>1.1713</td> <td>1.3978</td> <td>39</td> <td>1.334</td> </tr> <tr> <td>1.1764</td> <td>1.3997</td> <td>40</td> <td>1.375</td> </tr> <tr> <td>1.1816</td> <td>1.4016</td> <td>41</td> <td>1.415</td> </tr> <tr> <td>1.1868</td> <td>1.4036</td> <td>42</td> <td>1.456</td> </tr> <tr> <td>1.1920</td> <td>1.4056</td> <td>43</td> <td>1.498</td> </tr> <tr> <td>1.1972</td> <td>1.4076</td> <td>44</td> <td>1.539</td> </tr> <tr> <td>1.2025</td> <td>1.4096</td> <td>45</td> <td>1.581</td> </tr> <tr> <td>1.2296</td> <td>1.4200</td> <td>50</td> <td>1.796</td> </tr> <tr> <td>1.2575</td> <td>1.4307</td> <td>55</td> <td>2.020</td> </tr> <tr> <td>1.2865</td> <td>1.4418</td> <td>60</td> <td>2.255</td> </tr> <tr> <td>1.3163</td> <td>1.4532</td> <td>65</td> <td>2.500</td> </tr> </tbody> </table> </center>
<center><span>Table 1 Densities of cell structures in sucrose</span></center><center></center><center></center><center> <table> <tbody> <tr> <td>Structure</td> <td>Density (g/cm<sup>3</sup>)</td> </tr> <tr> <td>Golgi apparatus</td> <td>1.06-1.10</td> </tr> <tr> <td>Plasma membranes</td> <td>1.16</td> </tr> <tr> <td>Smooth endoplasmic reticulum</td> <td>1.16</td> </tr> <tr> <td>Intact oncogenic viruses</td> <td>1.16-1.18</td> </tr> <tr> <td>Mitochondria</td> <td>1.19</td> </tr> <tr> <td>Lysosomes</td> <td>1.21</td> </tr> <tr> <td>Peroxisomes</td> <td>1.23</td> </tr> <tr> <td>Plant viruses</td> <td>1.30-1.45</td> </tr> <tr> <td>Soluble proteins</td> <td>1.30</td> </tr> <tr> <td>Rhino- and enteroviruses</td> <td>1.30-1.45</td> </tr> <tr> <td>Nucleic acids, ribosomes</td> <td>1.60-1.75</td> </tr> <tr> <td>Glycogen</td> <td>1.70</td> </tr> </tbody> </table> </center>
