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Human PDE1B Insect Cell Lysate (WB positive control)

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Human PDE1B Transfected / Overexpression Cell Lysate Product Information
Expressed Host:Baculovirus-Insect cells
Product Description:Baculovirus-Insect Cell lysate that Human PDE1B transfected / overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).
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.
Predicted N Terminal:Met
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 Transfected / Overexpression Cell Lysate Usage Guide
Preparation Method:Cell lysate was prepared by homogenization 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.
Quality Control Testing:12.5% SDS-PAGE Stained with Coomassie Blue after protein purification.
Stability:Samples are stable for up to twelve months from date of receipt.
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. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
Storage Buffer:1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Storage Instruction:Store at 4℃. After re-dissolution, aliquot and store at -80℃.
Application notes:Western blot (WB): Use at an assay dependent dilution.
Other Applications: Not tested.
Optimal dilutions/concentrations should be determined by the end user.
PDE1B Background

Calcium/calmodulin-dependent 3',5'-cyclic nucleotide phosphodiesterase 1B, also known as Cam-PDE 1B and PDE1B, is a cytoplasm protein which belongs to the cyclic nucleotide phosphodiesterase family and PDE1 subfamily. Phosphodiesterase-10A (PDE10A), 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.

Human PDE1B 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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    Catalog: 11746-H20BL-300
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