| Product Name | SensoLyte ® MG Phosphate Assay Kit *Colorimetric* |
| Size | 1 kit |
| Catalog # | AS-71103 |
| US$ | $300 |
The MG (Malachite Green) Phosphate Assay Kit is based on quantification of the green complex formed between Malachite Green, molybdate, and free orthophosphate. The rapid color formation from the reaction can be conveniently measured on a spectrophotometer (600-660 nm) or on a plate reader. The non-radioactive colorimetric assay kits have been optimized to offer superior sensitivity and prolonged shelf life. The assay is simple and fast, involving a single addition step for phosphate determination. Assays can be performed in tubes, cuvettes, or multi-well plates. The assays can be conveniently performed in 96 or 384-well plates for high-throughput screening of enzyme inhibitors. The kit has been used for assaying phosphatase, phospholipase or lipase, nucleoside triphosphatase (ATPase, GTPase, etc.) and phytase. The kit has the followingcomponents: Highly purified Malachite Green reagent Protein phosphatase inhibitor Assay buffer A "mix and read" assay protocol that is compatible with HTS liquid handling instruments with Z" factors of 0.7 to 0.9. Kit size: 1000 assays | |
| Detailed Information | |
| Storage | Room temperature |
| References | Ref: Gurpal S. et al. (2003) Characterization of organic phosphorus in leachate from a grassland soil. Soil Biol.ogy & Biochemistry 35, 1317-1323; Cogan EB (1999) A robotics-based automated assay for inorganic and organic phosphates. Anal Biochem. 271, 29-35; Fisher DK and Higgins TJ (1994) A sensitive, high-volume, colorimetric assay for protein phosphatases. Pharm Res.11, 759-63; Henkel RD (1988) A microassay for ATPase. Anal Biochem. 169, 312-318. |
| Molecular Weight | N/A |
| Product Citations | Hwang, IW. et al. (2014). Human acetyl-CoA carboxylase 2 expressed in silkworm Bombyx mori exhibits posttranslational biotinylation and phosphorylation. Appl Microbiol Biotechnol 1.Lahiri, T. et al. (2014). A structural basis for the regulation of an H-NOX-associated cyclic-di-GMP synthase/phosphodiesterase enzyme by nitric oxide-bound H-NOX. Biochem 53, 2126.Maione, F. et al. (2014). Interleukin-17A exacerbates ferric chloride-induced arterial thrombosis in rat carotid artery. Intl J Inflamm, 2014. doi: http://dx.doi.org/10.1155/2014/247503.Day, S. et al. (2013). Impaired assembly and post-translational regulation of 26S proteasome in human end-stage heart failure. Circ Heart Fail 6, 544. doi: 10.1161/CIRCHEARTFAILURE.112.000119.Lu, D & PA. Insel (2013). Hydrolysis of extracellular ATP by ectonucleoside triphosphate diphosphohydrolase (ENTPD) establishes the set point for fibrotic activity of cardiac fibroblasts. J Biol Chem 288, 19040Midde, K. et al. (2013). Comparison of orientation and rotational motion of skeletal muscle cross-bridges containing phosphorylated and dephosphorylated myosin regulatory light chain. J Biol Chem 288, 7012. doi: 10.1074/jbc.M112.434209.Zaparaty, LM. et al. (2013). The first prokaryotic trehalose synthase complex identified in the hyperthermophilic Crenarchaeon Thermoproteus tenax. PlosOne. doi: 10.1371/journal.pone.0061354.Manque, PA. et al. (2012). Identification and characterization of a novel calcium-activated apyrase from Cryptosporidium parasites and its potential role in pathogenesis. PLoS 7, e31030. doi:10.1371/journal.pone.0031030Repen, B. et al. (2012). Optimization of a malachite green assay for detection of ATP hydrolysis by solubilized membrane proteins. Anal Biochem http://dx.doi.org/10.1016/j.ab.2012.04.016Clayton, A. et al. (2011). Cancer Exosomes Express CD39 and CD73, Which Suppress T Cells through Adenosine Production. J Immunol 187, 676. doi: 10.4049/jimmunol.1003884.Hilaire, C. et al. (2011). NT5E mutations and arterial calcifications. N Engl J Med 364, 432.Oomah, BD. et al. (2011). Phenolics, phytic acid, and phytase in Canadian-grown low-tannin Faba Bean (Vicia faba L.) genotypes. J Agric Food Chem 10.1021/jf200338b.Pinust, F. et al. (2011). Purification, primary structure, and properties of Euphorbia characias latex purple acid phosphatase. Biochem 76, 694.Yang, Z. et al. (2011). Characterization of LipL as a non-heme, Fe(II)-dependent α-ketoglutarate:UMP dioxygenase that generates uridine-5"-aldehyde during A-90289 biosynthesis. J Biol Chem 10.1074/jbc.M110.203562.Vickers, K. et al. (2010). Lyso-phosphatidylcholine induces osteogenic gene expression and phenotype in vascular smooth muscle cells. Atherosclerosis 211, 122.Wolf, A. et al. (2010). Plakophilin 1 stimulates translation by promoting eIF4A1 activity. JCB 188, 463. |
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做一次5-RACE的话,克隆10个以上基因应该是可以的。本数据来源于百度地图,最终结果以百度地图最新数据为准。
请采纳。。。。
我也想问一下。
步骤:1,RNA抽提(同RT)
2,逆转录,使用合成的oligo
3,p内参
4,使用根据已知序列设计的上游引物和提供下游引物做PCR,注意设计上游引物是退火温度与下游引物相差不大。
5,产物切胶回收,TA克隆测序
这个oligo和下游引物是我根据Takara和BD公司的manual自己设计的,经过我的使用效果很好。具体操作参照takara的3-racemanual。你先做3-race,了解熟悉下,成功后再做5-race。5-race要买试剂盒做,较为复杂!

