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蛋白提取及WB
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===转膜=== <ol type="1"> <li>转膜缓冲液的配制: <table class="wikitable" style="margin: auto; border-collapse: collapse; border: 1px solid #a2a9b1;"> <tr> <th colspan="2">1.5L 转膜缓冲液组成</th> </tr> <tr> <th>Reagent</th> <th>Quantity (for 1.5 L)</th> </tr> <tr style="border-top: 1px solid #a2a9b1;"> <td>Tris</td> <td>4.545 g</td> </tr> <tr> <td>甘氨酸</td> <td>21.6 g</td> </tr> <tr> <td>UP水</td> <td>1.2 L</td> </tr> <tr> <td>乙醇</td> <td>300 mL*</td> </tr> <tr style="border-top: 1px solid #a2a9b1;"> <td colspan="2"><i>*:乙醇待水溶液体系完全溶解后再加入</i></td> </tr> </table> <li>PVDF膜及其支撑膜浸入乙醇中活化5 min;转入转膜缓冲液浸泡;可在膜右上角剪口分辨方向 <li>转膜缓冲液倒入盘中,各层预先在盘中浸透 <li>黑色在下,按照海绵(层数可增加,能夹紧即可)-1层滤纸-gel(切割周围)-膜-1层滤纸-海绵,进行叠加,注意覆盖转膜液排出气泡,夹紧,按照标识放入电泳槽 <li>4 ℃冰水浴,300 mAh,电泳约50 min </ol>
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