磷酸二酯酶PDE IV底物 FAM-cAMP 绿色荧光

磷酸二酯酶PDE IV底物 FAM-cAMP 绿色荧光 是美国AAT Bioquest生产的荧光底物，这种绿色荧光cAMP衍生物是磷酸二酯酶（PDE）IV的特定底物。它可用于检测PDE IV活性或与FRET读数或FP格式的抗cAMP抗体一起筛选PDE IV抑制剂。 PDE是一组降解第二种信使分子的酶：环状核苷酸cAMP和cGMP。它们调节亚细胞域内环状核苷酸信号传导的定位，持续时间和幅度。因此，PDE是这些第二信使分子介导的信号转导的重要调节剂。 PDE酶由于其独特的组织分布，结构和功能特性，经常成为药理抑制的靶标。 PDE抑制剂可通过抑制PDE的降解来延长或增强cAMP或cGMP介导的生理过程的作用。在诸如肺动脉高压，冠心病，痴呆，抑郁症和精神分裂症等领域，PDE抑制剂被确定为新的潜在剂。，为您提供优质的磷酸二酯酶PDE IV底物 FAM-cAMP 绿色荧光 。 

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适用仪器

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荧光酶标仪
Ex:	490nm
Em:	525nm
Cutoff:	515nm
推荐孔板:	纯黑色孔板

样品实验方案 

溶液制备 

1.储备溶液

所有未使用的储备溶液应分为一次性使用的等分a试样，并在制备后储存在-20°C下。 避免重复冻融循环。
1.1 FAM-cAMP PDE IV储备溶液（1 mM）：
将500 µL DMSO加入0.5 umol FAM-cAMP PDE IV底物瓶中，制成1 mM储备液。

 

2.工作溶液

FAM-Cyclic-3'，5'-AMP PDE IV底物测定溶液（2X）：
通过将1 mM FAM-Cyclic-3'，5'-AMP PDE IV底物储备溶液稀释到您的PDE缓冲液（例如10 mM Tris-HCl）中，制成2X FAM-Cyclic-3'，5'-AMP PDE IV底物测定溶液 ，pH 7.4、10 mM的Mg Cl2、1 mM的MnCl2）制成200-400 nM的溶液。 注意：仅进行测定所需的足够数量。

 

样品操作及分析

1.将等体积的PDE IV标准品或样品与2X FAM-Cyclic-3'，5'-AMP PDE IV底物测定溶液混合，并在室温下孵育至少1小时。

2.在Ex / Em = 490/525 nm处监测荧光偏振。

 

参考文献

cAMP is a ligand for the tandem GAF domain of human phosphodiesterase 10 and cGMP for the tandem GAF domain of phosphodiesterase 11
Authors: Gross-Langenhoff M, Hofbauer K, Weber J, Schultz A, Schultz JE.
Journal: J Biol Chem (2006): 2841

cAMP phosphodiesterase activity evaluation in human carcinoma of salivary glands
Authors: Spoto G, della Malva M, Rubini C, Fioroni M, Piattelli A, Serra E, Di Nicola M, Santoleri F.
Journal: Nucleosides Nucleotides Nucleic Acids (2006): 1113

cAMP phosphodiesterase-4A1 (PDE4A1) has provided the paradigm for the intracellular targeting of phosphodiesterases, a process that underpins compartmentalized cAMP signalling
Authors: Huston E, Houslay TM, Baillie GS, Houslay MD.
Journal: Biochem Soc Trans (2006): 504

Characterization of a novel cAMP-binding, cAMP-specific cyclic nucleotide phosphodiesterase (TcrPDEB1) from Trypanosoma cruzi
Authors: Diaz-Benjumea R, Laxman S, Hinds TR, Beavo JA, Rascon A.
Journal: Biochem J (2006): 305

Compartmentalization of cAMP-Dependent Signaling by Phosphodiesterase-4D Is Involved in the Regulation of Vasopressin-Mediated Water Reabsorption in Renal Principal Cells
Authors: Stefan E, Wiesner B, Baillie GS, Mollajew R, Henn V, Lorenz D, Furkert J, Santamaria K, Nedvetsky P, Hundsrucker C, Beyermann M, Krause E, Pohl P, Gall I, Macintyre AN, Bachmann S, Houslay MD, Rosenthal W, Klussmann E.
Journal: J Am Soc Nephrol. (2006)

DdPDE4, a novel cAMP-specific phosphodiesterase at the surface of dictyostelium cells
Authors: Bader S, Kortholt A, Snippe H, Van Haastert PJ.
Journal: J Biol Chem (2006): 20018

Expression and Activity of cAMP Phosphodiesterase Isoforms in Pulmonary Artery Smooth Muscle Cells from Patients with Pulmonary Hypertension: Role for PDE1
Authors: Murray F, Patel HH, Suda RY, Zhang S, Thistlethwaite P, Yuan JX, Insel PA.
Journal: Am J Physiol Lung Cell Mol Physiol. (2006)

Expression of cAMP and cGMP-phosphodiesterase isoenzymes 3, 4, and 5 in the human clitoris: immunohistochemical and molecular biology study
Authors: Oelke M, Hedlund P, Albrecht K, Ellinghaus P, Stief CG, Jonas U, Andersson KE, Uckert S.
Journal: Urology (2006): 1111

Expression of cAMP and cGMP-phosphodiesterase isoenzymes 3, 4, and 5 in the human clitoris: immunohistochemical and molecular biology study
Authors: Sampaio F.
Journal: Int Braz J Urol (2006): 483

Helix-1 of the cAMP-specific phosphodiesterase PDE4A1 regulates its phospholipase-D-dependent redistribution in response to release of Ca2+
Authors: Huston E, Gall I, Houslay TM, Houslay MD.
Journal: J Cell Sci (2006): 3799

 

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