• 我要登录|
  • 免费注册
    |
  • 我的丁香通
    • 企业机构:
    • 成为企业机构
    • 个人用户:
    • 个人中心
  • 移动端
    移动端
丁香通 logo丁香实验_LOGO
搜实验

    大家都在搜

      大家都在搜

        0 人通过求购买到了急需的产品
        免费发布求购
        发布求购
        点赞
        收藏
        wx-share
        分享

        Preparation of Affinity Column制备亲和层析柱【UCSF】

        互联网

        1271

        1.Add 222.2μl 1M MOPS,pH 7.5 to 2ml of 10mg of CREBtide (0.1M MOPS pH 7.5 final concentration).

        2.Read OD205,OD280 (using 0.1M MOPS buffer as blank)

        3.Affigel-10 is stored frozen at < -70℃.Thaw at 4℃ for ~20min.

        4.Filter 1-2ml on nylon (0.22uM poresize),using Buchner funnel and vacuum.Do not allow beads to dry.

        5.Wash bed with 3x2ml dH2O.

        6.Scrape 0.5-1ml into 15ml microfuge tube.

        7.Add 2.22ml CREBtide ligand solution to tube.

        Rock at 4℃ for 4hrs.

        8.Transfer slurry to column.

        9.Wash 2x500μl dH2O.

        10.Collect eluate and save at -20℃.Read OD.

        11.Wash column:

        2x5ml cold PBS

        1x5ml PBS/0.02% NaN3 (1μl NaN3 in5ml PBS)

        12.Store at 4℃ upright in beaker in PBS/NaN3 parafilm.

        ad image
        提问
        扫一扫
        丁香实验小程序二维码
        实验小助手
        丁香实验公众号二维码
        扫码领资料
        反馈
        TOP
        打开小程序