| 查看: 1816 | 回复: 4 | |||
[交流]
求助酶活测定方法 PAL、CHS、F3H、DFR、LDOX
|
|
谁能提供以下植物中 酶活性测定方法： PAL、CHS、F3H、DFR、LDOX 谢谢了！ |
回复此楼» 猜你喜欢
医学类期刊求推荐
已经有5人回复
-
生活琐事由它去
已经有4人回复
-
提交了我也来说说感想
已经有12人回复
-
青B发送上会通知了吗
已经有9人回复
-
西安交大新媒学院副院长用撤稿论文结题
已经有6人回复
-
论文撤稿了
已经有8人回复
-
化学专业申博
已经有4人回复
-
某211大学教师把个人教师官方主页改成:我跑了我跑了我跑了!官宣跑路！
已经有5人回复
-
26/27申博自荐
已经有9人回复
-
博士申请
已经有3人回复
» 抢金币啦！回帖就可以得到:
-
坐标北京，诚征女友
+2/400
-
山东第一医科大学泰山学者团队2026年招收副教授、博士后（不限专业）
+1/80
-
【急招】 北京工业大学 第二批博士申请-杰青课题组1-2名额，25日截止！！
+1/40
-
双一流大学湘潭大学“化工过程模拟与强化”国家地方联合工程研究中心招收博士生
+1/34
-
南京农业大学工学院博士招生 1个名额
+1/28
-
郑州大学·安徽高等研究院 2026年博士招生（申请考核制）
+1/20
-
海南医科大学肖华教授(国家级青年人才)招收博士生1名---报名从速
+4/16
-
澳大利亚科廷大学（Curtin University）招收全奖博士生
+1/9
-
【有偿访谈招募】高才通来港后，你过得还好吗？
+1/7
-
杭州师范大学博士后招聘
+1/6
-
【有偿访谈招募】高才通来港后，你过得还好吗？
+1/6
-
紧急招收2026年秋季入学博士生1名（河北工大/北京科技大学联合 增材制造/生物材料）
+1/5
-
汕头大学医学院黄乃淇教授实验室招聘博士后
+1/5
-
【有偿访谈招募】高才通来港后，你过得还好吗？
+1/4
-
【有偿访谈招募】高才通来港后，你过得还好吗？
+1/4
-
【有偿访谈招募】高才通来港后，你过得还好吗？
+1/4
-
求助专利一篇，谢谢
+1/3
-
北航杭州国际校区招聘3D 打印、陶瓷材料等博后
+1/3
-
易度质量流量计在防水透声膜气体透过率测试中的应用
+1/2
-
博士去绵阳华丰科技怎么样，待遇如何，这个企业靠谱吗？
+3/1
2楼2014-09-15 14:32:02
★
小木虫: 金币+0.5, 给个红包，谢谢回帖
小木虫: 金币+0.5, 给个红包，谢谢回帖
|
这个好像很难，我查到了2种方法 第一种：ASSAY OF CHALCONE SYNTHASE ACTIVITY BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY，PhytochemistVryo,l . 34,N o. 5, pp. 1225-1229，1993 第二种： CHS[12] chaleone synthase，查儿酮合成酶 Chemicals, [2-14C]Malonyl-CoA (1.71GBq•mmo1-1) was purchased from N.E.N. (Boston, Mass., USA) and [U-14C]phenyl-alanine (16.65GBq•mmol-1) from Amersham (Braunschweig, Germany). 4-Coumaroyl-CoA was synthesized by Dr. P.A. Bäumker (Institut für Botanik, Münster, Germany; Bäumker et al. 1988)[13]. Enzyme preparations. ö Pbenylalanine ammonia-lyase: Leaves were cut into small pieces, frozen at -20°C and homogenized in a prechilled mortar in cold acetone (- 20°C The homogenate was filtered and the residue washed several times with cold acetone. The resulting dry powder was suspended in 0.1 M sodium borate buffer (pH 8.8) containing 5 mM dithioerythritol (DTE). Chalcone synthase. All steps were carried out at 0-4°C Leaf material was homogenized in 0.1 M potassium phosphate buffer (pH 6.8) containing 40 mM ascorbate, 1 mM EDTA, 10 mM thiourea硫脲, 5 mM DTE and 0.25 g Polyclar AT.(g FW)-1 (Serva, Heidelberg, Germany). The homogenate was centrifuged for 10 min at 10 000 g and the supernatant concentrated by ultrafiltration (Diaflo PM 30; Amicon, Witten, Germany). Enzyme assays, The activity of PAL was measured by the method of Amrhein and Zenk (1971). The activity of CHS was determined according to Beerhues and Wiermann (1988)[14], except that the pH of the incubation buffer was 7.8 and the final DTE concentration 0.5 mM. The analysis of the reaction products was performed as described by Lembach et al. (1989)[15]. (CHS activity was determined according to Beerhues and Wiermann (1985)[16]. The analysis of the reaction product was carried out using thin layer chromatography with silica gel plates and hexane: ethyl-acetate: methanol = 50:50:l (v/v/v) as a solvent system. For re-chromatography of the eluted product cellulose plates and 15% (v/v) ethanol were employed.) Protein determination. Protein concentration was measured according to the method of Bradford (1976), as modified by Read and Northcote (1981), using ovalbumin as a standard. Schöpker, H., M. Kneisel, L. Beerhues, et al. Phenylalanine ammonia-lyase and chalcone synthase in glands of Primula kewensis (W. Wats): immunofluorescence and immunogold localization. Planta, 1995. 196(4): 712-719. 13. PA, B., A. S and W. R. Metabolism of ferulic acid sucrose esters in anthers of Tulipa cv. Apeldoorn. II. Highly specific degradation of the esters by different esterase activities. Z. Naturforsch, 1988. 43c: 647-655. |
3楼2014-12-26 14:49:21
4楼2017-10-18 21:48:59
5楼2018-08-05 13:29:58