Human PDE3A Baculovirus-Insect cells Overexpression Lysate

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Human PDE3A Baculovirus-Insect cells Overexpression Lysate: 产品信息

产品描述
This Human PDE3A overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of PDE3A protein (Cat: 11908-H20B1) from the overexpression lysate was verified.
表达宿主
Baculovirus-Insect cells
种属
Human
蛋白构建信息
A DNA sequence encoding the human PDE3A (Q14432) (Lys669-Gln1141) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
分子量
The recombinant human PDE3A /GST chimera consists of 710 amino acids and has a calculated molecular mass of 81.7 kDa. The recombinant protein migrates as an approximately 93 kDa band in SDS-PAGE under reducing conditions.

Human PDE3A Baculovirus-Insect cells Overexpression Lysate: 使用指南

制备方法
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
裂解缓冲液
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
使用建议
1.  Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2.  Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
缓冲液
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
应用
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human PDE3A Baculovirus-Insect cells Overexpression Lysate: 别称

Human CGI-PDE Overexpression Lysate; Human CGI-PDE-A Overexpression Lysate; Human CGI-PDEA Overexpression Lysate

PDE3A 背景信息

PDE3A belongs to the phosphodiesterase family. Phosphodiesterases (PDEs) are a family of related phosphohydrolyases that selectively catalyze the hydrolysis of 3' cyclic phosphate bonds in adenosine and/or guanine 3',5' cyclic monophosphate (cAMP and/or cGMP). They regulate the cellular levels, localization and duration of action of these second messengers by controlling the rate of their degradation. PDEs are expressed ubiquitously, with each subtype having a specific tissue distribution. These enzymes are involved in many signal transduction pathways and their functions include vascular smooth muscle proliferation and contraction, cardiac contractility, platelet aggregation, hormone secretion, immune cell activation, and they are involved in learning and memory. PDE3A has a high affinity for both cAMP and cGMP and shows competitive inhibition of the cAMP hydrolytic activity by cGMP. It plays a critical role in regulating intracellular levels of cAMP and cGMP.
全称
phosphodiesterase 3A, cGMP-inhibited
参考文献
  • Penmatsa H, et al. (2010) Compartmentalized cyclic adenosine 3',5'-monophosphate at the plasma membrane clusters PDE3A and cystic fibrosis transmembrane conductance regulator into microdomains. Mol Biol Cell. 21(6):1097-110.
  • Zhang W, et al. (2007) Thrombin regulates intracellular cyclic AMP concentration in human platelets through phosphorylation/activation of phosphodiesterase 3A. Blood. 110(5):1475-82.
  • Degerman E, et al. (1997) Structure, localization, and regulation of cGMP-inhibited phosphodiesterase (PDE3). J Biol Chem. 272(11):6823-6.
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