Méchin Valérie, Damerval Catherine, Zivy Michel
UMR de Chimie Biologique, Thiverval Grignon, France.
Methods Mol Biol. 2007;355:1-8. doi: 10.1385/1-59745-227-0:1.
We describe a procedure allowing extraction of total proteins that performs efficiently with a large variety of plant tissues, based on simultaneous precipitation and denaturation with TCA and 2ME in cold acetone. We also describe protein solubilization prior to IEF, either in classical rod gels or in IPGs, using two different solutions. The procedure is easy to carry out. The major caveats are (1) keep samples at low temperature during extraction, and then (2) manage protein samples at about 22 to 25 degrees C to avoid urea precipitation.
我们描述了一种能从多种植物组织中高效提取总蛋白的方法,该方法基于在冷丙酮中用三氯乙酸(TCA)和2-巯基乙醇(2ME)同时进行沉淀和变性。我们还描述了在等电聚焦(IEF)之前,在经典柱状凝胶或固相pH梯度胶(IPG)中使用两种不同溶液溶解蛋白质的方法。该方法易于实施。主要注意事项为:(1)提取过程中将样品保持在低温状态,然后(2)在约22至25摄氏度下处理蛋白质样品以避免尿素沉淀。