Overview

  • Product nameAnti-cAMP antibody [SPM486]
    See all cAMP primary antibodies
  • Description
    Mouse monoclonal [SPM486] to cAMP
  • Tested applicationsSuitable for: Flow Cyt, ELISA, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    Adenosine 3, 5-Cyclic Monophosphate (cAMP) compounds.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage bufferpH: 7.40
    Preservative: 0.05% Sodium azide
    Constituents: BSA, 0.0268% PBS
  • Concentration information loading...
  • PurityProtein G purified
  • ClonalityMonoclonal
  • Clone numberSPM486
  • IsotypeIgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab24851 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
Flow Cyt Use at an assay dependent concentration. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ELISA Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration.

WB was tested against chemically linked cAMP-carrier protein which was used for antibody screening.

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 [SPM486] images

  • ab24851 staining cAMP in MALME-3M cells treated with melanotan II (ab141161), by ICC/IF. Increase of cAMP expression correlates with increased concentration of melanotan II, as described in literature.
    The cells were incubated at 37°C for 4h in media containing different concentrations of ab141161 (melanotan II) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature 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 ab24851 (5 μg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 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.
  • ICC/IF image of ab24851 stained HeLa 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 (ab24851, 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.

  • ab24851 staining cAMP in MALME-3M cells treated with α-MSH (ab120205), by ICC/IF. Increase of cAMP correlates with increased concentration of α-MSH, as described in literature.
    The cells were incubated at 37°C for 2h in media containing different concentrations of ab120205 (α-MSH) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature 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 ab24851 (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.
  • ab24851 staining cAMP in MALME-3M cells treated with α-MSH (ab120189), by ICC/IF. Increase of cAMP correlates with increased concentration of α-MSH as described in literature.
    The cells were incubated at 37°C for 6h in media containing different concentrations of ab120189 (α-MSH) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature 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 ab24851 (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 [SPM486] (ab24851)

This product has been referenced in:
  • Weber M  et al. Mechanisms of cyclic nucleotide phosphodiesterases in modulating T cell responses in murine graft-versus-host disease. PLoS One 8:e58110 (2013). Flow Cyt ; Mouse . Read more (PubMed: 23483980) »
  • Vaeth M  et al. Dependence on nuclear factor of activated T-cells (NFAT) levels discriminates conventional T cells from Foxp3+ regulatory T cells. Proc Natl Acad Sci U S A 109:16258-63 (2012). Flow Cyt . Read more (PubMed: 22991461) »

See all 5 Publications for this product

Product Wall


To answer your questions:

1. No, the antibody is not conjugated.

2. You can use an anti-mouse secondary antibody. For PE conjugation we have a couple:

ab97041 (http://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-M...

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ab24851 is not conjugated therefore, you would need a secondary anti-mouse IgG conjugated to a fluorophore. I could suggest ab7003 conjugated to PE (http://www.abcam.com/ab7003) or ab150111 conjugated to Alexa 647(http://www.abcam.com/ab150111). Howe...

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I am sorry for the delay in getting back to you.

I now have further information to share with you in regards to the carrier protein used and the size od band that this has shown in Western blotting. The cAMP compounds used in Western blotting ...

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Thank you very much for that information.

I am sorry but I can still not share any further information relating to the conjugation used with this antibody. I will get back to you as soon as I can with this information.

Thank you for t...

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Thank you for contacting us. I am sorry for the delay in getting back to you. My colleague who has been dealing with your case is currently away from the office but I will try to assist as best I can.

My colleague has been in contact with the ...

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Thank you for contacting Abcam.

I am sorry about the issues you were having with ab24851. As we discussed on the telephone, I am sending you a new vial of ab24851 to try out in flow cytometry. Your new order number is **********. When you rec...

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Thank you for your email. I am sorry to hear that you are experiencing problems with this antibody.

I have checked the protocol which looks fine to however I have following suggestions and questions;

- IHc-Fr application gives best...

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Thank you for taking the time tocontact us. I am sorry to hearthe customer hashad difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this case for you and this is ...

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Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly. Having reviewe...

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Thank you for your enquiry. I have checked with the laboratory but unfortunately, it seems that we do not have the ELISA data for this product. If you would like the general protocol that we use for testing in ELISA, please refer to the protocol pag...

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