Human PDE1B Baculovirus-Insect cells Overexpression Lysate

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Human PDE1B Baculovirus-Insect cells Overexpression Lysate: Product Information

Product Description
This Human PDE1B overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of PDE1B protein (Cat: 11746-H20B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Species
Human
Sequence Information
A DNA sequence encoding the human PDE1B long isoform (Q01064-1) (Met 1-Asp 536) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus.
Molecule Mass
The recombinant human PDE1B/GST chimera consists of 773 amino acids and has a calculated molecular mass of 89.2 kDa. It migrates as an approximately 75 kDa band in SDS-PAGE under reducing conditions.

Human PDE1B Baculovirus-Insect cells Overexpression Lysate: Usage Guide

Preparation Method
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.
Lysis Buffer
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.
Recommend Usage
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.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
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.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human PDE1B Baculovirus-Insect cells Overexpression Lysate: Alternative Names

Human PDE1B1 Overexpression Lysate; Human PDES1B Overexpression Lysate

PDE1B Background Information

Calcium/calmodulin-dependent 3',5'-cyclic nucleotide phosphodiesterase 1B, also known as Cam-PDE 1B and PDE1B, is a cytoplasm protein which belongs to thecyclic nucleotide phosphodiesterase family and PDE1 subfamily. Phosphodiesterase-1A (PDE1A), Phosphodiesterase-1B (PDE1B), Phosphodiesterase-4B (PDE4B), and Phosphodiesterase-4A (PDE4A) are important regulators of signal transduction in striatum due to their catalysis of cyclic AMP and cyclic GMP. PDE1B is highly expressed in the striatum. It binds two divalent metal cations per subunit. Site one of PDE1B may preferentially bind zinc ions, while site two of PDE1B has a preference for magnesium and/or manganese ions. PDE1B is a cyclic nucleotide phosphodiesterase with a dual-specificity for the second messengers cAMP and cGMP, which are key regulators of many important physiological processes. It has a preference for cGMP as a substrate.
Full Name
phosphodiesterase 1B, calmodulin-dependent
References
  • Reed,T.M. et al.,1998, Mamm Genome. 9 (7):571-6.
  • Zhang K.Y.J. et al., 2004, Mol. Cell 15:279-86.
  • Siuciak,J.A. et al., 2007, Neuropharmacology. 53 (1):113-24.
  • Dlaboga, D. et al., 2008, Neuropharmacology. 54 (4):745-54.
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