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5 L of 1/100 dilution of the Alexa Fluor 647-anti cAMP antibody solution in the Detection Buffer (PerkinElmer Life and Analytical Sciences) and 10 L of 1/5000 dilution of Eu-W8044 labeled streptavidin and Biotin-cAMP in the Detection Buffer (PerkinElmer Life and Analytical Sciences) were added in reduced lighting conditions