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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

今日热帖
  • 精华评论
  • 菲菲菲鱼

    引用回帖:
    8楼: Originally posted by iknow at 2018-12-19 13:13:59
    已发送。主要查收

    好的,谢谢

  • 胡哲flight

    可以测到赤霉素和油菜素内酯吗

  • iknow

    引用回帖:
    10楼: Originally posted by 胡哲flight at 2018-12-29 14:56:28
    可以测到赤霉素和油菜素内酯吗

    可以测到赤霉素GA1,3和4. 其他赤霉素形式没有标准品。油菜素内脂也没有标准品。如果你有标准品,我可以测试一下

  • mazhq1988

    引用回帖:
    10楼: Originally posted by 胡哲flight at 2018-12-29 14:56:28
    可以测到赤霉素和油菜素内酯吗

    得分材料,我目前都测出来过

  • iknow

    引用回帖:
    12楼: Originally posted by mazhq1988 at 2019-01-21 11:28:58
    得分材料,我目前都测出来过...

    恩。我们没有油菜素内脂标准品。所以没有建立方法。

  • 大王椰子

    能发一下王学敏2008年的文献吗,谢谢,892526283@qq.com

  • kanhong

    王学敏的论文名称方便发一下么?谢谢楼主

  • flyever2012

    引用回帖:
    15楼: Originally posted by kanhong at 2019-08-15 12:10:08
    王学敏的论文名称方便发一下么?谢谢楼主

    同求此文献,如果你有的话,可以发我一份吗

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