cAMP Cell-Based Assays

Cisbio Bioassays offers a line of three ready-to-use assays for measuring cAMP using HTRF® technology. Each kit is optimized for detecting a specific range of cAMP concentrations to allow fine tuning of assay sensitivity. These kits enable high quality, cell-based assay screening in 96-, 384- or 1536-well formats.

Features:
  • Streamlined single-plate protocol for HTS
  • Low false positive, false negative results
  • Stable EC50 from 30 minutes to 7 days
  • Miniaturization down to <10µl
  • Compatible with fresh or frozen cells
Applications:
  • Gs or Gi coupled receptor activation
  • Agonist / antagonist screening
  • Phosphodiesterase (PDE) monitoring
  • Adenylate cyclase activity assessment

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Assay Principle:

 

cAMP kits are based on a competitive immunoassay using cryptate-labeled anti-cAMP and d2-labeled cAMP. d2 is our newest HTRF acceptor fluorophore whose small size eliminates EC50 drift.
To also satisfy all the robotics constraints, each kits are supplied with a 1 or 2 dispensing steps protocol after cell stimulation



Product Specifications:

 

Three kit groups to cover a broad range of cell-based assays.

With the introduction of Lumi4™-Tb cryptate, cAMP femto, the most sensitive cAMP assay is now available with two cryptates (europium or terbium). Both kits enable the detection of as little as 28 pM cAMP (0.6 fmoles/well in 20µl volume). This allows use of very low cell densities, 100 to 500 cells per well.

The cAMP dynamic 2 - wide range of cAMP concentrations ideal for a broad range of Gi/s assays.

The cAMP HiRange - well suited for HTS with extended signal-to-background ratios (Z' > 0.9) and for use with membrane preparations.

  Cryptate S/B EC20 nM* EC50 nM* EC80 nM*
cAMP femto 2 Kit Eu3+ 10 0.33 1.76 9.4
cAMP femto Tb Kit Tb2+ 10 0.33 1.76 9.4
cAMP dynamic 2 Kit Eu3+ 18 0.91 4.07 18.1
cAMP HiRange Kit Eu3+ 41 4.27 22.8 121
* final cAMP concentration



Assay Performance:

 

All HTRF cAMP kits are unaffected by culture media additions such as serum, biotin, or colored compounds and tolerate > 5% DMSO. cAMP assays have been validated using fresh suspension cells, adherent cells and frozen cells. The plates were read at several intervals following cell stimulation, from 1 hour to 7 days. The IC50 values calculated with the three kits are remarkably stable even after 7 days, which is a significant advantage during screening campaigns.

CHO-K1 cells were stimulated with Methylhistamine in the presence of 30 µM forskolin (EC80). 20 µl assays were performed in 384-well plates. Plates were read at times ranging from 1 hour to 7 days.

Ordering Info:

 

cAMP Cell-based Assay Reagents:

Description Quantity* Cat no. Product Insert
cAMP femto 2 kit 1,000 tests 62AM5PEB pdf
20,000 tests 62AM5PEC pdf
100,000 tests 62AM5PEJ pdf
cAMP femto Tb 1,000 tests 62AM7PEB pdf
20,000 tests 62AM7PEC pdf
100,000 tests 62AM7PEJ -
cAMP dynamic 2 kit 1,000 tests 62AM4PEB pdf
20,000 tests 62AM4PEC pdf
100,000 tests 62AM4PEJ pdf
cAMP HiRange kit 1,000 tests 62AM6PEB pdf
20,000 tests 62AM6PEC pdf
100,000 tests 62AM6PEJ pdf
* based on 20 µl assay volume

Companion Products:

Description Cat no.
cAMP femto 2 calibrator 62AM5CDA
cAMP femto 2 control 62AM5TDA
cAMP dynamic 2 calibrator 62AM4CDA
cAMP dynamic 2 control 62AM4TDA
cAMP HiRange calibrator 62AM6CDA
cAMP HiRange control 62AM6TDA
Diluent for standard curve prep. 62DL1DDD


For more information or To Place an Order:
 

U.S. and Canada
1-888-963-4567

Find your local sales and support team

Europe and other countries
+33(0)466 796 705

Japan
+81-3-5510-2932