Overview

  • Product name
    cAMP Assay Kit - Direct Immunoassay
    See all cAMP kits
  • Sample type
    Urine, Serum, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media
  • Assay type
    Quantitative
  • Sensitivity
    > 0.02 µM
  • Range
    0.02 µM - 2 µM
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Mammals
  • Product overview

    cAMP Assay Kit (ab65355) provides a direct competitive immunoassay for sensitive and quantitative determination of cAMP level in biological samples. The kit utilizes recombinant Protein G coated 96-well plate to efficiently anchor cAMP polyclonal antibody on to the plate. cAMP-HRP conjugate directly competes with cAMP from sample binding to the cAMP antibody on the plate. After incubation and washing, the amount of cAMP-HRP bound to the plate can easily be determined by reading HRP activity at OD450 nm. The intensity of OD450 nm is inversely proportional to the concentration of cAMP in samples. In addition, the kit provides a new acetylation procedure to improve detection sensitivity significantly. The kit can detect ~0.1-10 pmol/5 µl (or ~ 0.02-2 µM) cAMP samples.
    Visit our FAQs page for tips and troubleshooting.

  • Notes

    Adenosine 3’, 5’ cyclic monophosphate (cAMP) is an important second messenger in intracellular signal transduction. Monitoring cAMP levels is one of the most common ways to screen for agonists and antagonists of GPCRs.

     

    If you are using fluorometric detection, we recommend cAMP Direct Immunoassay Detection Kit (Fluorometric) (ab138880).

     

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Storage buffer
    Preservative: None
  • Components Identifier 100 tests
    10X cAMP Assay Buffer WM 1 x 25ml
    Acetylating Reagent A Violet 1 x 750µl
    Acetylating Reagent B Black 1 x 1.5ml
    cAMP-HRP Green 1 vial
    HRP Developer Amber 1 x 10ml
    Neutralizing Buffer NM 1 x 7.5ml
    Protein G Coated Plate 1 unit
    Rabbit Anti-cAMP polyclonal Antibody Red 1 vial
    Standard cAMP (10 nmol) Yellow 1 x 10µl
  • Research areas
  • Relevance
    Adenosine 3’,5’-cyclic monophosphate (cyclic AMP, cAMP) is an important “second messenger” involved in many physiological processes. It plays a key role as an intracellular second messenger for transduction events that follow a number of extracellular signals.
  • Alternative names
    • Adenosine 3’ 5’ cyclic monophosphate
    • Cyclic adenosine monophosphate
    • Cyclic AMP

Images

  • Measurement of cAMP in Jurkat and HepG2 cells with ab65355. Assay performed using the kit protocol.

  • Standard curve (colorimetric): mean of duplicates (+/-SD)

Protocols

References

This product has been referenced in:
  • Wu L  et al. C1QTNF1 attenuates angiotensin II-induced cardiac hypertrophy via activation of the AMPKa pathway. Free Radic Biol Med 121:215-230 (2018). Read more (PubMed: 29733904) »
  • Liu Z  et al. CgA1AR-1 acts as an alpha-1 adrenergic receptor in oyster Crassostrea gigas mediating both cellular and humoral immune response. Fish Shellfish Immunol 58:50-58 (2016). Read more (PubMed: 27633678) »

See all 15 Publications for this product

Customer reviews and Q&As

As this kit will detect a cyclic nucleotide and not a protein, it does not have a species restriction and should be fine for the use with bacteria.

We recommend starting with 10 Million cells or 100 mg of tissue.

A possibility is that esterase degraded the cAMP. So preserve your sample fresh, and use 0.1M HCl lyse your sample immediately. cAMP is stable in 0.1M HCl.

You are correct. There is no need to add a blocking step to this protocol. Further you do not need to wash the plates prior to adding the antibody or cAMP-HRP.

The kit was easier to use and worked as expected. Directions for sample prep were followed and we had measurable values of cAMP on this assay. The protocols are fairly simple but required thorough review to understand certain steps. Our sample pool in serial dilution performed similar to the standards, however need to be diluted less than the kit recommends for our sample type. We will continue using this assay in the future.
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Abcam user community

Verified customer

Submitted Sep 21 2017

Most labs generally have a bench-top vacuum centrifuge which I think would be ideal for this purpose, but if not, the oven drying should work as well. Use an oven at ˜60oC.

The cAMP antibody affinity is higher than that of cGMP. That is the reason why you need fmol of cAMP while for cGMP you need pmol quantities.

I am pleased to provide the suggested protocol for preparing cell lysate samples for this assay:

Cell Samples:

Aspirate medium.
Add 1 ml of 0.1M HCl for every 35 cm2 of surface area.
Incubate at room temperature for 20 m...

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The hemoglobin will not bind to the antibody in the plate. It will go out with the wash.

In regards to your question as to whether ourcAMP Direct Immunoassay Kit, ab65355, will work for your need, yes this assay can work beautifully for such a purpose. Please use all appropriate background controls.

This product is covered by ou...

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