• 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

        Crystallization Trials

        互联网

        917

        Materials

        Milipore filter type HA 0.45 micron
        Culture plates (Linbro model 76-033-05)
        Protein in DDW or HEPES buffer (10 mg/mL)
        Vacuum grease
        Syringe with 18 G needle

        Procedure

        1. Millipore filter all solutions to be used.
        2. Dialyze protein for 24 hours against water, if possible. Otherwise, include minimum requirements for keeping the protein soluble. If buffer is necessary use HEPES buffer.
        3. For the initial screening use the buffers and precipitants recommended in the fast crystal screen (Jancarik, J. & Kim, S.H. 1991 J. Appl. Cryst. 24, 409), as instructed , and the hanging drop method with the Linbro tissue culture plates that have 24 wells, 1.7 cm diameter, 1.6 cm deep. See diagram .
        4. Use a protein solution with a starting concentration of 10 mg/mL. This concentration may have to be adjusted depending on the number of the drops that precipitate. The concentration is about correct when one-third to one-half of the drops develop precipitate within one week.
        5. Deposit 0.7 mL of reservoir solution into the well. Combine 2 microliters of the protein solution and 2 microliters of the reservoir solution to make up the protein droplet.
        6. Incubate one at room temperature and one at cold (4-10°C) temperature.

        Notes

        1. Crstallization screening kits and other supplies are available from Hampton Research .

        Taken from methods by J. Jancarik and S.H. Kim

        If you use methods from the Kinesin site, we ask that you cite the Kinesin Home Page and authors, or the appropriate source publication in your work.

         

        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