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植物丝裂原活化蛋白激酶OsMAPK6(mitogen-activated protein kinase)活性检测ELISA 试剂盒 【植物OsMAPK6 酶联免疫试剂盒检测原理】本实验采用双抗体夹心ELISA 法,参考Yu et al.(2001)和 Goettert et al. (2012)所描述的方法,并进行改进。所提供的ELISA Kit 是典型的夹心法酶联免疫吸附测定试剂盒(ELISA)。Anti-OsMAPK6(又名Anti-OsMAPK1;北京华大蛋白质研发中心有限公司,产品编号:AbP80140-A-SE)是OsMAPK6磷酸化特异性抗体。预先包被的是待测OsMAPK6,能特异性识别具有活性的OsMAPK6,但不能识别非磷酸化不具活性的OsMAPK6。预先在96管中包被的是OsMAPK6。检测相抗体经生物素(biotin)标记。样品和生物素标记抗体先后加入酶标板孔反应后,PBS 或TBS 洗涤。随后加入过氧化物酶标记的亲和素反应;经过PBS 或TBS 的彻底洗涤后用底物TMB 显色。TMB 在过氧化物酶的催化下转化成蓝色,并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的待测OsMAPK6活性呈正相关。  Description of OsMAPK6 (mitogen-activated protein kinase) activity assay ELISA kit This mitogen-activated protein kinase activity analysis ELISA kit was developed by the methods described by Yu et al.(2001) and Goettert et al. (2012),with improvement. As a typical sandwich ELISA kit, it is less time-consuming, easy to perform, having good sensitivity and specificity to detect the activity of OsMAPK6. Anti-OsMAPK6 is a phospho-specific antibody against OsMAPK6. Anti-OsMAPK6 can recognize specifically the phosphorylated/activated but not the non-phosphorylated/inactive OsMAPK6.Anti-OsMAPK6 was purchased from Beijing Protein Innovation(also named anti-OsMPK1, Cat No. Abp80140-A-SE ). A standard curve of OsMAPK6 was made with different concentrations of 200, 100, 50, 25, 12.5, 6.25 and 3.12 (U/mL).The sample of OsMAPK6 was added and pre-coated to the 96-well plante and incubated at 37 °C for 90 min. After washing the plates twice with by TBS solution, 100 μL of HRP-conjugated anti-OsMAPK6 (Beijing protein innovation) was added and incubated at 37 °C for 60 min. Subsequently, 100 μL TMB color solution was added at 37 °C for 10 min. At last 100 μL stop solution (sulfuric acid) was added and the OD (450nm) was measured with EnSpire Multimode Reader (PerkinElmer, America).The activity of OsMAPK6 is positively correlated with the the depth of colour detected. 参考文献(References) Yu JS , Chang SH , Chan WH, et al. Enzyme-Linked Immunosorbent Assay for the Determination of p21-Activated Kinase Activity[J]. Journal of Biochemistry, 2001(2):243-251.  Goettert M , Shaalan N , Graeser R , et al. Development of a p38δ mitogen activated protein kinase ELISA assay for the quantitative determination of inhibitor activity.[J]. J Pharm Biomed Anal, 2012, 66(none):349-351.

产品类型:

国产/进口

产品名称:

植物丝裂原活化蛋白激酶OsMAPK6活性检测ELISA试剂

英文名称:

(Name of product):OsMAPK6 (mitogen-activated protein kinase) activity assay ELISA kit

货号:

(CAT No.):2Pl-KMLJ92048p-DZ

执行标准:

(Executive standard):HY03/J19

规格:

(Type):96T

中文价格:

2500元

种属:

(Species):Rice

待测物名称:

(Type of product):液体类物质 liquit

缩写:

(abbreviation):OsMAPK6

蛋白功能1:

(Item 1 of detection ):size

蛋白功能3:

( Item 3 of detection ):Mitogen-activated protein kinase activity assay

检测范围:

(Range):200U/ml-3.12U/ml

灵敏度:

(Sensitivity):0.6U/ml.

反应时间:

4-5h

所需样本体积:

100ul

检测波长:

(Detection wave length):450nm

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