This assay kit is a competition enzyme-linked immunoassay designed to measure cAMP levels in cells or tissues. An anti-mouse IgG was pre-coated onto a microplate. The cAMP in the test sample competes with HRP-cAMP for binding to an anti-cAMP mouse monoclonal antibody. Following the incubation and wash step, an ultra-sensitive HRP fluorescence substrate is added to generate fluorescence can be measured at excitation 535 nm and emission at 585 nm. Because of the competitive nature of this assay, the fluorescent intensity is inversely proportional to the quantity of sample cAMP. Measurement using the cAMP Standard allows calculating the absolute amount of cAMP in a sample. This assay is more sensitive than a colorimetric assay.
Typical Data
This standard curve is provided for demonstration only. A standard curve should be generated for each set of samples assayed. Fig. 2 is an example of typical Data.
Sensitivity
The minimum detectable dose (MOD) of cAMP is typically 1 nM.
The Intra-assay CV and the Inter-assay CV are ˂10%.
Specificity
This assay recognizes cAMP. No cross-reactivity with others.
Standard Preparation
Standard Curve
Kit Content and Storage Conditions
Part
Part#
Description
Storage of Opened/Reconstituted
Precoated Microplate
TBS2081A
96 well polystyrene microplate (12 strips of 8 wells) coated with anti-mouse IgG antibody.
Return unused wells to the foil pouch. Reseal along the entire edge of the zip-seal. May be stored for up to 1 month at 2-8 °C.
cAMP Standard
TBS2081B
50 µL of cAMP (1 mM).
Aliquot and store at -20 ℃ for up to 1 month in a manual defrost freezer. Avoid repeated freeze-thaw cycles.
Detection A
TBS2081C
100 µL of cAMP-HRP (25x).
Store at 4℃ for 3 months
Detection B
TBS2081D
1 mL of anti-cAMP antibody (5x).
Store at -20℃ for 12 months.
Assay Diluent
TBS2081E
15 mL of a buffered protein base with preservatives.
Store at 4℃ for 12 months.
HRP Fluorescent Substrate
TBS2081F
12 mL of ultra-sensitive fluorescent substrate.
Store at -20℃ for 12 months.
Substrate Developer
TBS2081G
20 µL
Store at 4℃ for 12 months.
Wash Buffer
TBS3000W
12 mL of concentrated solution (10x)
Store the unopened kit at 2-8 ℃. Do not use past kit expiration dates.
The kit contains sufficient materials to run an ELISA on one 96-well plate.
