Overview

  • Product name

    cAMP Assay Kit (Competitive ELISA)
    See all cAMP kits
  • Detection method

    Colorimetric
  • Sample type

    Saliva, Serum, Tissue Extracts, Cell Lysate, Cell culture media
  • Assay type

    Competitive
  • Assay time

    3h 00m
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Other species, Mammals
  • Product overview

    cAMP ELISA Kit ab133051 uses a sensitive colorimetric competitive ELISA method to quantify cAMP (cyclic AMP) levels in samples from mammals and other species. It has a maximum sensitivity of 0.039 pmol/mL / 0.039 nM. It is not suitable for use with samples from rabbits.


    The cAMP ELISA uses a 96-well plate supplied coated with a goat anti-rabbit IgG antibody. The anti-rabbit antibody binds an anti-cAMP antibody which is added to the plate.


    Alkaline phosphatase (AP)-labeled cAMP, and free cAMP within a sample, are then added to each well of the plate. The cAMP ELISA is based on the competition between the AP-labeled cAMP and the free cAMP for the fixed number of cAMP antibody binding sites on the plate.


    The more free cAMP is present, the less AP-labeled cAMP is bound. After a washing step, the amount of bound AP-labeled cAMP is detected using the colorimetric AP substrate pNpp measured by absorbance at OD 405 nm. The level of signal is compared to a standard curve for free cAMP, which is run at the same time as the samples.


    This kit uses an optional acetylation step to achieve the maximum sensitivity of 0.039 nM, without the acetylation step the sensitivity is 0.3 pmol/mL / 0.3 nM.

  • Notes

    This kit was previously called Cyclic AMP Complete ELISA Kit.

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

     

    Cross Reactivity 

    Compound Cross Reactivity
    cAMP 100%
    AMP <0.33%
    ATP <0.12%
    cGMP <0.001%
    GMP <0.001%
    GTP <0.001%
    cUMP <0.001%
    CTP <0.001%
  • Platform

    Microplate

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 1 x 96 tests
    0.1M HCl 1 x 27ml
    20X Wash Buffer Concentrate 1 x 27ml
    Acetylation kit - Acetic Anhydride 1 x 1ml
    Acetylation kit - Triethylamine 1 x 2ml
    Assay Buffer 2 1 x 27ml
    Cyclic AMP Alkaline Phosphatase Conjugate 1 x 5ml
    Cyclic AMP Complete Antibody 1 x 5ml
    Cyclic AMP Complete Standard 1 x 500µl
    Goat anti-rabbit IgG Microplate (12 x 8 wells) 1 unit
    Neutralizing Reagent 1 x 5ml
    Plate Sealer 1 unit
    pNpp Substrate 1 x 20ml
    Stop Solution 1 x 5ml
  • Research areas

  • Relevance

    dblinks not applicable
  • Alternative names

    • 3'-5'-cyclic adenosine monophosphate
    • Cyclic adenosine monophosphate
    • cyclic AMP

Images

  • Representative Standard Curve using ab133051. Non-Acetylated Format.

  • Representative Standard Curve using ab133051. Acetylated Format.

Protocols

References

This product has been referenced in:

  • Lari E  et al. Oil sands process-affected water impairs the olfactory system of rainbow trout (Oncorhynchus mykiss). Ecotoxicol Environ Saf 170:62-67 (2019). Read more (PubMed: 30529621) »
  • Zamarbide M  et al. Male-Specific cAMP Signaling in the Hippocampus Controls Spatial Memory Deficits in a Mouse Model of Autism and Intellectual Disability. Biol Psychiatry 85:760-768 (2019). Read more (PubMed: 30732858) »
See all 4 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Abreviews
Pig plasma from pigs treated with the drug PTH(1-34) was used in this experiment. The drug PTH(1-34) is reported to increase plasma cAMP levels. Pig plasma samples were diluted 10-fold with assay buffer 2. all other steps were done according to the protocol of the ab133051 ELISA Kit.

Abcam user community

Verified customer

Submitted Aug 04 2017

Murine Splenocyte ELISA

Excellent Excellent 5/5 (Ease of Use)
Abreviews
This ELISA worked well for detecting cAMP in total splenocytes from a C57BL/6J mouse. Cells were stimulated with 5uM IBMX and 5uM forskolin in a 96 well plate at a concentration of 2 million cells per well for 30 mins or left unstimulated. The cells were lysed in 20uls of 0.1M HCL per well.

Chris Wilson

Verified customer

Submitted Jan 12 2017

Answer




The “regular” cAMP kit, Cyclic AMP ELISA Kit (ab133039) comes with an assay buffer for diluting standards and samples, and is intended for use with extracellular cAMP – samples like serum or cell culture media, for example.


In the “direct” cAMP kit., Cyclic AMP Direct ELISA Kit (ab133038), the assay buffer has been replaced with 0.1M HCl and a neutralizing reagent has been included. Samples such as cell culture cells or tissue can be lysed in that HCl, then loaded directly into the wells (to which the neutralizing reagent has been added), hence the “direct” in the name.


The “complete” kit, Cyclic AMP Complete ELISA Kit (ab133051) simply includes both the assay buffer from the regular kit and the HCl/ neutralizing reagent combo from the direct kit so you can measure samples from anything. Please note that all the other components of the kit – plate, antibody, conjugate, standard, etc – are identical in all three cases.


As you would like to measure cell lysates and supernatant samples and therefore I would recommend kit ab133051.


Please note that the sensitivity is always higher when the samples is acetylated.


Read More

Cyclic AMP in mouse Bone marrow derived macrophages

Excellent Excellent 5/5 (Ease of Use)
Abreviews
We used this kit to measure cAMP in Bone marrow derived macrophages ( BMDMs) from mouse. Samples were collected in 0.1 M HCL and kept at -20ºc until use ( 2 weeks later). We used the acetylated format and the procedure was really easy. We can confirm with our data that the kit works perfectly in BMDMs from mouse.

Abcam user community

Verified customer

Submitted Jul 22 2015

Answer

Thank you for your enquiry.
I appreciate your patience, I have been discussing this question with the laboratories.

Actually all three cAMP kits are essentially the exact same kit; all kit components are identical with the exception of the buffers/materials provided to process the samples.

The ab133038 kit is intended for intracellular cAMP detection and comes with 0.1MHCl for lysis and sample/standard dilution as well as a neutralization reagent to balance the pH.
The ab133039 kit is intended for extracellular cAMP detection and comes with assay buffer for sample/standard dilution.
The complete cAMP kit, ab133039, simply comes with 0.1MHCl, neutralization reagent, and assay buffer so that researchers can look at both intracellular and extracellular cAMP.

I hope this will be helpful. If you have any further questions, please do not hesitate to contact me.

Read More

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