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: Mammal
  • 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).

     

  • Tested applications
    Suitable for: Functional Studiesmore details

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

Applications

Our Abpromise guarantee covers the use of ab65355 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Functional Studies Use at an assay dependent dilution.

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:
  • 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) »
  • Pan B  et al. Unique Effects of Acute Aripiprazole Treatment on the Dopamine D2 Receptor Downstream cAMP-PKA and Akt-GSK3ß Signalling Pathways in Rats. PLoS One 10:e0132722 (2015). Functional Studies ; Rat . Read more (PubMed: 26162083) »

See all 14 Publications for this product

Customer reviews and Q&As

Thank you for contacting us regarding this protocol.

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.

I hope that this info...

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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



I contacted the laboratories and they suggest that 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.

I


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.
However, we have not tested this ourselves, and therefore, cannot send you a protocol accordin...

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The species is not applicable for this kit. The species reactivity information we have on the datasheet is to show the kit was tested with human samples.

cAMP is a nucleotide which should be same in all species so this kit should work for thi...

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Thank you for your enquiry.

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. Read More

Thank you for your reply.

It is difficult to know what happened with the kit without extra protocol and sample information. Perhaps you can find some helpful information in our ELISA resources pages:

http://www.abcam.com/index.html?...

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Thank you for contacting us.





In regards to your question as to whether ourcAMP Direct Immunoassay Kit, ab65355, will work for your need, yesthis assay can work beautifully for such a purpose. Please use all appropriat...

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Thank you for contacting us.

cAMP Direct Immunoassay Kit utilizes recombinant Protein G coated 96-well plate to anchor cAMP polyclonal antibody on to the plate.

I have read through the protocol and couldn’t find the need for a...

Read More

Thank you for contacting us.

The standard concentration is indeed 2 pm/ul when diluted with 800ul of 0.1M HCL.

Step d (page 9) add 200ul of standard into 10pmol/50ul labelled vial, then we recommend doing the serial dilution. We t...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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