Overview

  • Product nameAnti-cAMP antibody [M486]
    See all cAMP primary antibodies
  • Description
    Mouse monoclonal [M486] to cAMP
  • Tested applicationsSuitable for: IHC-Fr, ICC/IF, WB, Indirect ELISAmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    A chemically linked 3', 5'-Cyclic Adenosine Monophosphate (cAMP).

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferPreservative: None
    Constituents: PBS, pH 7.2 containing antibody stabilizer
  • Concentration information loading...
  • PurityAscites
  • ClonalityMonoclonal
  • Clone numberM486
  • IsotypeIgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab70280 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
IHC-Fr Use at an assay dependent concentration. PubMed: 22649251
ICC/IF Use at an assay dependent concentration.
WB Use a concentration of 0.1 - 1 µg/ml. Predicted molecular weight: 1 kDa. WB was tested against chemically linked cAMP-carrier protein which was used for antibody screening.
Indirect ELISA Use a concentration of 0.01 - 0.1 µg/ml.

Target

  • RelevanceCyclic adenosine monophosphate (cAMP) plays a key role as an intracellular second messenger for transduction events that follow a number of extracellular signals. The G-Protein Coupled Receptors (GPCR) is the largest family of cell surface receptors. They can be activated by different ligands, such as neurotransmitters, hormones, ions, small molecules, peptides, and other physiological signaling molecules. Typically, the binding of the ligands to its receptor resulting in the activation of G-proteins, in return, activates the effector adenylyl cyclase evoking the production of cAMP. The activation of a protein kinase by cAMP results in the phosphorylation of substrate proteins. Currently successful drugs in marketing have been developed to target these receptors. Among the GPCRs, ~367 receptors are potential drug development targets, but only about 20 have been used to generate therapeutically and commercially successful drugs so far. Because the involvement of cAMP can amplify the response of the ligand binding, the second messenger cAMP has been largely employed to monitor the activation of the GPCR to facilitate the therapeutic drug discovery.
  • Cellular localizationSecreted
  • Alternative names
    • 3' 5' cyclic adenosine monophosphate antibody
    • Cyclic adenosine monophosphate antibody
    • Cyclic AMP antibody

Anti-cAMP antibody [M486] images

  • ELISA Plot:. Adenosine-3', 5'-cyclic AMP immobilized onto plates, followed by addition of stand cyclic AMP. The mouse anti c-AMP was added subsequentially, and visiualized by chromatogenic substrate. Each sample was done in triplicate. IC50 was then calculated.
  • ICC/IF image of ab70280 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab70280, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • ab70280 staining cAMP in HEK293 cells treated with neuropeptide S (ab120174), by ICC/IF. Increase in cAMP expression correlates with increased concentration of neuropeptide S, as described in literature.
    The cells were incubated at 37°C for 10 minutes in media containing different concentrations of ab120174 (neuropeptide S) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab70280 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
  • ab70280 staining cAMP in HEK293 cells treated with neuropeptide S (ab120246), by ICC/IF. Increase in cAMP expression correlates with increased concentration of neuropeptide S, as described in literature.
    The cells were incubated at 37°C for 10 minutes in media containing different concentrations of ab120246 (neuropeptide S) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab70280 (5 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.

References for Anti-cAMP antibody [M486] (ab70280)

This product has been referenced in:
  • Corredor RG  et al. Soluble adenylyl cyclase activity is necessary for retinal ganglion cell survival and axon growth. J Neurosci 32:7734-44 (2012). IHC-Fr ; Rat . Read more (PubMed: 22649251) »
  • Ogata G  et al. Dopamine and background illumination activate D1 and D2-D5-type receptors in adult rat retinal ganglion cells. J Comp Neurol : (2012). Rat . Read more (PubMed: 22678972) »

See all 2 Publications for this product

Product Wall

Thank you very much for contacting us with your question.

Using ab70280, the following protocol has given good results in ICC/IF:

1) Fix cells with 4% formaldehyde for 10 minutes
2) Permeablize and block the cells by incubating...

Read More

Thank you for your patience.

It is unclear what the journal means by "exact structure" if the table that is referred to in the instructions is meant to contain text. The text description of the immunogen is

Adenosine 3', 5' cyclic...

Read More

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