|Product Includes||Quantity (with Count)||Solution Color|
|cAMP Rabbit mAb Coated Microwells||1 x 96 tests|
|cAMP-HRP Conjugate||1 x 5.5 ml||Red|
|cAMP Standard (2.4 uM)||1 x 0.5 ml|
|Luminol/Enhancer Solution||1 x 3 ml|
|Stable Peroxide Buffer||1 x 3 ml|
|Sealing Tape||1 x 2 ea|
|ELISA Wash Buffer (20X) 9801||1 x 10 ml|
|Cell Lysis Buffer (10X) 9803||1 x 15 ml|
NOTE: If cell debris is observed it can be removed by brief centrifugation of the plate and transfer of the clear lysates to a new 96 well plate.
Make cAMP standard in the 1X Cell Lysis buffer: Take 50 µl of the cAMP standard (2.4 µM) and add it to 450 µl diluent to get 240 nM cAMP. Perform a 1:3 serial dilution of this standard to get 80 nM, 26.7 nM, 8.9 nM, 3.0 nM, 1.0 nM, 0.3 nM,and 0 nM. The diluent without cAMP will serve as the 0 nM cAMP.
NOTE: The standard curve is used to calculate the absolute amount of cAMP in the sample and is necessary for each assay.
Use a plate-based luminometer to measure Relative Light Units (RLU) at 425nM within 1-10 minutes following addition of the substrate. Optimal signal intensity is achieved when read within 10 minutes.
Protocol Id: 506
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