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植物激素提取和测定方法交流-Elisa和质谱方法

作者 iknow
来源: 小木虫 1150 23 举报帖子
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最近在摸索的植物激素提取和测定方法,主要是采用液质联用仪法。在论坛里面看来很多帖子,自己也查阅了很多文献,其中pp上有一篇文献multiple phytohormone profiling by targeted metabolomics中的方法,只需要用20mg新鲜组织就可以提取所有激素。但是需要用到层析柱。
具体方法供大家参考。:
For the quantifcation of targeted plant hormones and related compounds,
20-mg (fresh weight) portions of separately harvested roots and shoots were
weighed into 2-mL plastic microtubes (Eppendorf) and frozen in liquid nitrogen. To minimize the risk of false-positive detections, possible background
levels of analytes were subtracted from measured sample values. Therefore,
in the blank controls, 1 mL of extraction buffer containing a mixture of stable
isotopically labeled IS also was purifed. Before extraction, three 3-mm ceriastabilized zirconium oxide beads (Next Advance) were added to each sample. A mixture of stable, isotopically labeled IS was added to validate the
method and enable the precise quantifcation of endogenous levels of targetedcompounds. The amounts of IS ranged from 0.4 to 50 pmol per sample (precise amounts are listed in Supplemental Table S3). Frozen plant material was
extracted in 1 mL of ice-cold 50% aqueous (v/v) ACN using an MM 301vibration mill (Retsch) operating at a frequency of 27 Hz for 5 min. Samples were
then sonicated for 3 min at 4°C using a Transsonic T310 ultrasonicator with an
ice block-flled bathtub (Elma) and subsequently extracted using a Stuart SB3
benchtop laboratory rotator (Bibby Scientifc) for 30 min at 4°C. After centrifugation (10 min, 36,670 g, and 4°C; Beckman Avanti 30), the supernatant was
transferred to clean plastic microtubes. Samples were further purifed according to the scheme shown in Figure 1D.

我们目前参照王学敏2008年文献,不使用层析柱,也能测到大部分激素。

在做激素测到的同志可以留言一起交流交流。 返回小木虫查看更多

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  • 精华评论
  • 菲菲菲鱼

    没看见 Figure 1D么

  • iknow

    引用回帖:
    2楼: Originally posted by 菲菲菲鱼 at 2018-12-19 08:50:57
    没看见 Figure 1D么

    可以查原始文献。multiple phytohormone profiling by targeted metabolomics

  • iknow

    如果无法下载原始文献,可以加我qq,我发给你。或者把邮箱留下,我发你邮箱。

  • 菲菲菲鱼

    引用回帖:
    4楼: Originally posted by iknow at 2018-12-19 09:47:59
    如果无法下载原始文献,可以加我qq,我发给你。或者把邮箱留下,我发你邮箱。

    发我邮箱吧,1401751838@qq.com

  • iknow

    引用回帖:
    5楼: Originally posted by 菲菲菲鱼 at 2018-12-19 10:17:45
    发我邮箱吧,1401751838@qq.com...

    好的。

  • 菲菲菲鱼

    引用回帖:
    6楼: Originally posted by iknow at 2018-12-19 11:49:26
    好的。...

    谢谢

  • iknow

    引用回帖:
    7楼: Originally posted by 菲菲菲鱼 at 2018-12-19 12:21:39
    谢谢

    已发送。主要查收

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