Adenylate Cyclase 1 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.

Adenylyl cyclases function to convert ATP to cyclic AMP in response to activation by a variety of hormones, neurotransmitters and other regulatory molecules. Cyclic AMP, in turn, activates several other target molecules to control a broad range of diverse phenomena such as metabolism, gene transcription and memory. Adenylyl cyclases respond to receptor-initiated signals, mediated by the Gs and Gi heterotrimeric G proteins. The binding of an agonist to a Gs-coupled receptor catalyzes the exchange of GDP (bound to Galpha s) for GTP, the dissociation of GTP-Galpha s from Gbetagamma and Galpha s)-mediated activation of adenylyl cyclase. Adenylyl cyclases type I (AC I) and III (AC III) have distinct staining within the cell nucleus of rat brain sensory neurons. AC I is expressed in myenteric ganglia as two bands of 160 kDa and 185 kDa by SDS-PAGE. Ca2+ stimulation of AC I and AC III is mediated by calmodulin. Protein associated with Myc (PAM) is a very potent inhibitor of AC I. A decrease in endogenous PAM levels in HeLa cells modulate both basal and agonist stimulated cAMP accumulation.

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 NBP2-06564

• Adenylate Cyclase 1 Antibodies
• Adenylate Cyclase 1 ELISA Kit
• Adenylate Cyclase 1 Lysate