<track id="j575v"><span id="j575v"><th id="j575v"></th></span></track>
<address id="j575v"></address>

    <video id="j575v"><progress id="j575v"><progress id="j575v"></progress></progress></video>

      <p id="j575v"></p>

      <form id="j575v"><dl id="j575v"></dl></form>
      <video id="j575v"></video><nobr id="j575v"></nobr><video id="j575v"></video>
        <track id="j575v"></track>

        <pre id="j575v"></pre><big id="j575v"><span id="j575v"></span></big>

            您當前所在位置: 首頁 > 學者


            • 75瀏覽

            • 0點贊

            • 0收藏

            • 0分享

            • 58下載

            • 0評論

            • 引用


            Improvement of gel-separated protein identification by DMF-assisted digesion and peptide recovery after electroblotting

            王賢純 , Yong Lin , Yan Li , Wenjun Han , Quanze He , Jianglin Li , Ping Chen , Xianchun Wang , Songping Liang*

            Electrophoresis 2009, 30, 3626-3635,-0001,():



            In-gel digestion of gel-separated proteins is a major route to assist in proteomics-based biological discovery, which, however, is often embarrassed by its inherent limitations such as the low digestion efficiency and the low recovery of proteolytic peptides. For overcoming these limitations, many efforts have been directed at developing altemative methods to avoid the in-digestion. Here, we present a new method for efficient protein digestion and tryptic peptide recoery, which involved electroblotting gel-separated proteins onto a PVDFmembrane, excising the PVDF bands containing protein of interest, and dissolving the bands with pure DMF (≥99.8%). Before tryptic digestion, NH4HCO3 buffer was added to moderately adjust the DMF concentration (to 40%) in order for trypsin to exert its activity. Experimental results using protein standards showed that, due to actions of DMF in dissolving PVDF membrane and the membrane-bound substances, the proteins were virtually insolution digested in DMF-containing buffer. This protocol allowed more efficient digestion and peptide recovery, thereby in creasing the sequence coverage and the confidence of protein identification. The comparative study using rat hippocampal membrane-enriched sample showed that the method was superior to the reported on-membrane tryptic digestion for further protein identification, including low abundant and/or highly hydrophobic membrane proteins.

            【免責聲明】以下全部內容由[王賢純]上傳于[2009年11月13日 10時43分04秒],版權歸原創者所有。本文僅代表作者本人觀點,與本網站無關。本網站對文中陳述、觀點判斷保持中立,不對所包含內容的準確性、可靠性或完整性提供任何明示或暗示的保證。請讀者僅作參考,并請自行承擔全部責任。


            全部評論 0