PDE4B (Rat) Assay Kit

Phosphodiesterases (PDEs) play an important role in the dynamic regulation of the second messengers cAMP (cyclic adenosine monophosphate) and cGMP (cyclic guanosine monophosphate) signaling, by hydrolyzing them. The PDE superfamily is composed of 11 families, with PDE4,7 and 8 being cAMP-specific hydrolases, and thus regulating positive and negative responses to it. PDE4 is the second most abundantPDEin cardiovascular tissues. PDE4, is a cAMP-specific PDE and the largest PDE subfamily with over 35 differentisoformsand as such, the most widely characterized PDEisoenzyme. Inhibitors of PDE4 are being used for the treatment of psoriasis, COPD (chronic obstructive pulmonary disease) and atopic dermatitis. PDE4B is the most abundant PDE4 in the brain and it is involved in cognitive function and memory, and diseases such as schizophrenia and neuroinflammation. Evidence points for the formation of a PDE4B/DISC1 (Disrupted in Schizophrenia 1) complex that regulates PKA (Protein kinase A) phosphorylation of NDEL1 (nuclear distribution factor-E-like) and its interaction with LIS1 (lissencephaly 1), and thus impacts synaptic function. In the context of neuroinflammation PDE4B is linked to the NF-κB (necrosis factor kappa B) signaling pathway via the regulation of cAMP levels in cells. High levels of PDE4B have also been found in colon cancer and hematological disorders such as CLL (chronic lymphocytic leukemia) . The development of PDED4B specific inhibitors and a deeper understanding of the role of each PDE4B isoform in health and disease will prove beneficial for treatment of PDE4B related disorders.

Catalog #NameAmountStorage60049Rat PDE4B, GST-Tag*˃1 µg-80°C60200FAM-Cyclic-3′, 5′-AMP**1.2 nmoles**-80°C60393PDE Assay Buffer (Incomplete) 25 ml-20°C60390PDE Binding Agent100 µl4°C60391Binding Agent Diluent (cAMP) 10 ml4°C827350.5 M DTT200 µl-20°C79685Low binding, black 96-well plate1Room Temp* The concentration of protein is lot-specific and will be indicated on the tube containing the protein.** FAM-Cyclic-3′, 5′-AMP is provided as a powder. Vial will need to be resuspended in 600 µl of Complete PDE Assay Buffer before use.

The final concentration of DMSO in the assay should not exceed 1%.Fluorescent compounds that have λex=470 nm and detection at λem=528 nm can interfere with the readouts.It is recommended that the compound alone is tested to determine any potential interference of the compound on the assay results.