PDE1B Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

PDE1B, a variant of PDE1, is a Ca2+-calmodulin-stimulated phosphodieterase (PDE) preferentially hydrolyzing cGMP. Cyclic nucleotides are hydrolyzed and compartmentalized by a family of enzymes called phosphodieterases. In mammals at least 11 different families of PDEs can be discriminated (PDE1-11) based on their kinetic properties and inhibition to various pharmacological agents. Three different PDE1 genes have been identified (PDE1A, PDE1B, PDE1C). The enzymes of the PDE1A and PDE1B have higher affinity for cGMP than for cAMP, whereas PDE1C has high affinity for both cAMP and cGMP. Calcium-Calmodulin stimulates the activity of each of these enzymes several-fold. The tissue distribution of PDE1B is different from other PDE1 gene family members. In testis PDE1B is expressed uniformly through out the seminiferous epithelium and in the interstistium in contrast to PDE1A and PDE1C that is expressed in elongated to mature spermatids.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBL1-14215

• PDE1B Antibodies
• PDE1B Lysate
• PDE1B Proteins