Overview

  • Product name

    Cyclic AMP Direct ELISA Kit
    See all cAMP kits
  • Detection method

    Colorimetric
  • Sample type

    Tissue, Cell Lysate
  • Assay type

    Competitive
  • Assay time

    3h 00m
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: a wide range of other species
  • Product overview

    Abcam’s Cyclic AMP in vitro competitive ELISA (Enzyme-Linked Immunosorbent Assay) is a colorimetric competitive immunoassay kit for the quantitative determination of intracellular cAMP in cells or tissues lysed in 0.1 M HCl.

    A goat anti-rabbit IgG antibody has been precoated onto 96-well plates. Standards or test samples are added to the wells, along with an alkaline phosphatase (AP) conjugated-cAMP antigen and a polyclonal rabbit antibody specific to cAMP. After incubation the excess reagents are washed away. pNpp substrate is added and after a short incubation the enzyme reaction is stopped and the yellow color generated is read at 405 nm. The intensity of the yellow coloration is inversely proportional to the amount of cAMP captured in the plate.

  • Notes

    Cyclic AMP (cAMP) is one of the most important “second messengers” involved as a modulator of physiological processes. A number of hormones are known to activate cAMP through the action of the enzyme Adenylate cyclase which converts ATP to cAMP. cAMP has been shown to be involved in the cardiovascular and nervous systems, immune mechanisms, cell growth and differentiation, and general metabolism. 

    Cross Reactivity

    Compound% Cross Reactivity
    cAMP100
    AMP0.33
    ATP0.12
    cGMP<0.001
    GMP<0.001
    GTP<0.001
    cUMP<0.001
    CTP<0.001
  • Tested applications

    Suitable for: Competitive ELISAmore details
  • 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
    cAMP Antibody 1 x 5ml
    Cyclic AMP Alkaline Phosphatase Conjugate 1 x 5ml
    Cyclic AMP Complete Standard 1 x 0.5ml
    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

Applications

Our Abpromise guarantee covers the use of ab133038 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
Competitive ELISA Use at an assay dependent concentration.

Images

  • Representative Standard Curve using ab133038

  • Representative Standard Curve using ab133038

Protocols

References

This product has been referenced in:

  • Moussa BA  et al. Synthesis and molecular docking of new roflumilast analogues as preferential-selective potent PDE-4B inhibitors with improved pharmacokinetic profile. Eur J Med Chem 148:477-486 (2018). Read more (PubMed: 29477888) »
  • Thouverey C & Caverzasio J Suppression of p38a MAPK Signaling in Osteoblast Lineage Cells Impairs Bone Anabolic Action of Parathyroid Hormone. J Bone Miner Res 31:985-93 (2016). Read more (PubMed: 26643857) »
See all 2 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

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

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

Answer

Gracias por contactarnos de nuevo, y por tu soporte a nuestros productos. Siento la tardanza en contestar.

Respecto a las preguntas del kit ab133038:

1) La fecha de caducidad del producto varía de lote a lote. Nosotros garantizamos su funcionamiento en los 6 meses siguientes a la fecha de compra, y siempre recomendamos comprar el kit cuando vaya a utilizarse, en lugar de dejarlo almacenado y usarlo al cabo de los meses.

2) Respecto a las publicaciones, me temo que si en la datasheet del producto no hay ninguna indicada, es porque lamentablemente no tenemos constancia de ninguna referencia en la que se haya usado este kit. Tan pronto como conozcamos de la existencia de alguna referencia trabajando con este kit, actualizaremos la página web.

3) El kit está pensado para la detección intracelular de cAMP. Siempre que haya cAMP presente en las muestras, el kit debiera detectarlo. No tenemos experiencia en el uso del kit en células de osteosarcoma de rata, pero recomendaría a la clienta que buscara literatura para conocer los niveles de cAMP esperados en las muestras de interés.

Espero que esta información sea útil para la clienta. En caso contrario, no dudes en comentármelo, y estaré encantada de poder ayudar.

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Answer

Thank you for your enquiry.

The kit is provided with break apart strips which can be used only once.

Just let me know if there is any more information you need.

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