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植物激素提取和测定方法交流-Elisa和质谱方法 已有11人参与
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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。 |
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