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cAMP(Cyclic adenosine monophosphate) ELISA Kit

Citations(52)

  • Cat.No.:E-EL-0056

  • Reactivity: Universal
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  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
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Product Details

Properties

Assay type Competitive-ELISA
Format 96T/48T
Assay time 2.0h
Detection range 1.56-100 ng/mL
Sensitivity 0.94 ng/mL
Sample type &Sample volume serum, plasma and other biological fluids; 50μL
Specificity This kit recognizes cAMP in samples. No significant cross-reactivity or interference between cAMP and analogues was observed.
Reproducibility Both intra-CV and inter-CV are < 10%.
Application This ELISA kit applies to the in vitro quantitative determination of cAMP concentrations in serum, plasma and other biological fluids.

Test Principle

This ELISA kit uses the Competitive-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with cAMP. During the reaction, cAMP in the sample or standard competes with a fixed amount of cAMP on the solid phase supporter for sites on the Biotinylated Detection Ab specific to cAMP. Excess conjugate and unbound sample or standard are washed away, and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well and incubated. Then a TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns from blue to yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The concentration of cAMP in tested samples can be calculated by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.
ItemSpecificationsStorage
Micro ELISA Plate(Dismountable) 96T: 8 wells ×12 strips
48T: 8 wells ×6 strips 		-20℃, 12 months
Reference Standard 96T: 2 vials
48T: 1 vial
Concentrated Biotinylated Detection Ab (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL 		-20℃(Protect from light), 12 months
Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°C, 12 months
Biotinylated Detection Ab Diluent 1 vial, 14 mL
HRP Conjugate Diluent 1 vial, 14 mL
Concentrated Wash Buffer (25×) 1 vial, 30 mL
Substrate Reagent 1 vial, 10 mL 2-8℃(Protect from light)
Stop Solution 1 vial, 10 mL 2-8℃
Plate Sealer 5 pieces
Manual 1 copy
Certificate of Analysis 1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level cAMP were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level cAMP were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (ng/mL) 5.40 14.10 37.40 5.90 15.00 34.90
Standard deviation 0.30 0.70 2.00 0.40 0.90 1.80
CV (%) 5.56 4.96 5.35 6.78 6.00 5.16

Recovery

The recovery of cAMP spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum(n=8) 89-104 95
EDTA plasma(n=8) 86-100 92
Cell culture media(n=8) 87-100 92

Linearity

Samples were spiked with high concentrations of cAMP and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

SynonymsCyclic adenosine monophosphate, cAMP, cyclic AMP, 3'-5'-cyclic adenosine monophosphate
Research AreaSignal transduction

Assay Procedures

elisa assay procedure 1

1. Add 50μL standard or sample to the wells, immediately add 50μL Biotinylated Detection Ab working solution to each well. Incubate for 45 min at 37°C
elisa assay procedure 2

2. Aspirate and wash the plate for 3 times
elisa assay procedure 3

3. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
elisa assay procedure 4

4. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
elisa assay procedure 5

5. Add 50μL Stop Solution
elisa assay procedure 6

6. Read the plate at 450nm immediately. Calculation of the results

Citations

  1. Journal of Ginseng Research (2022) IF: 6.06
    Ginsenoside Rg1 promotes neurite growth of retinal ganglion cells through cAMP/PKA/CREB pathways

    DOI: 10.1016/j.jgr.2022.05.002
    Sample: Rgc-5S Cell
  2. PROGRESS IN NEURO-PSYCHOPHARMACOLOGY & BIOLOGICAL PSYCHIATRY (2022) IF: 5.201
    Neuronal lack of PDE7a disrupted working memory, spatial learning, and memory but facilitated cued fear memory in mice

    DOI: 10.1016/j.pnpbp.2022.110655

    PMID: 36220621

    		 Sample: Hippocampus Samples
  3. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2022) IF: 5.924
    Receptor-Mediated AKT/PI3K Signalling and Behavioural Alterations in Zebrafish Larvae Reveal Association between Schizophrenia and Opioid Use Disorder

    DOI: 10.3390/ijms23094715

    PMID: 35563106

    		 Sample: Larvae
  4. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2022) IF: 5.924
    Pyruvate Upregulates Hepatic FGF21 Expression by Activating PDE and Inhibiting cAMP–Epac–CREB Signaling Pathway

    DOI: 10.3390/ijms23105490
    Sample: Hepg2 Cell
  5. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2018) IF: 5.924
    Developmental Exposure to Atrazine Impairs Spatial Memory and Downregulates the Hippocampal D1 Dopamine Receptor and cAMP-Dependent Signaling Pathway in Rats

    DOI: 10.3390/ijms19082241

    PMID: 30065202

    		 Sample: Tissue Homogenate
  6. Frontiers in Pharmacology (2022) IF: 5.988
    Inhibitory effect of (pro)renin receptor decoy inhibitor PRO20 on endoplasmic reticulum stress during cardiac remodeling

    DOI: 10.3389/fphar.2022.940365

    PMID: 36034809

    		 Sample: Cardiomyocytes (Cms),Cardiac Fibroblasts (Cfs)
  7. ANDROLOGIA (2019) IF: 1.84
    Effect of low androgen status on the expression of adenosine A2A and A2B receptors in rat penile corpus cavernosum

    DOI: 10.1111/and.13344
    Sample: Corpus Cavernosum
  8. BMC Medical Genetics (2018) IF: 1.913
    Identification of a novel GNAS mutation in a case of pseudohypoparathyroidism type 1A with normocalcemia

    DOI: 10.1186/s12881-018-0648-z

    PMID: 30060753

    		 Sample: Cell Culture Supernatant
  9. International Journal of Stem Cells (2020) IF: 1.94
    Estrogen Promotes cAMP Production in Mesenchymal Stem Cells by Regulating ADCY2

    DOI: 10.15283/ijsc19139

    PMID: 32114743

    		 Sample: Cell Culture Supernatant
  10. Experimental and Therapeutic Medicine (2018) IF: 2.447
    Calcimimetic R568 reduced the blood pressure and improved aortic remodeling in spontaneously hypertensive rats by inhibiting local renin‑angiotensin system activity

    DOI: 10.3892/etm.2018.6734
    Sample: Plasma,Tissue Homogenate
  11. Experimental and Therapeutic Medicine (2020) IF: 2.447
    Calcilytic NPS2143 promotes proliferation and inhibits apoptosis of spontaneously hypertensive rat vascular smooth muscle cells via activation of the renin‑angiotensin system

    DOI: 10.3892/etm.2020.8759
    Sample: Cell Lysate
  12. FITOTERAPIA (2020) IF: 2.527
    Anti-melanogenic effects of epigallocatechin-3-gallate (EGCG), epicatechin-3-gallate (ECG) and gallocatechin-3-gallate (GCG) via down-regulation of cAMP/CREB /MITF signaling pathway in B16F10 melanoma cells

    DOI: 10.1016/j.fitote.2020.104634

    PMID: 32454171

    		 Sample: Cell Culture Supernatant
  13. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS (2018) IF: 2.559
    rLj-RGD3, a novel recombinant toxin protein from Lampetra japonica, prevents coronary thrombosis-induced acute myocardial infarction by inhibiting platelet functions in rats

    DOI: 10.1016/j.bbrc.2018.02.021
    Sample: Platelets
  14. Reproductive Sciences (2020) IF: 2.616
    Quinoline Derivative Enhances Human Sperm Motility and Improves the Functional Competence

    DOI: 10.1007/s43032-020-00382-5

    PMID: 33237520

    		 Sample: Spermatozoa (Spz)
  15. Frontiers in Neurology (2020) IF: 2.889
    PKA-Dependent Membrane Surface Recruitment of CI-AMPARs Is Crucial for BCP-Mediated Protection Against Post-acute Ischemic Stroke Cognitive Impairment.

    DOI: 10.3389/fneur.2020.566067
    Sample: Tissue Homogenate
  16. JOURNAL OF CELLULAR BIOCHEMISTRY (2018) IF: 2.959
    A novel role and mechanism of cystic fibrosis transmembrane conductance regulator in bisphenol A‐induced prostate cancer

    DOI: 10.1002/jcb.28156
    Sample: Cell Culture Supernatant
  17. EXPERIMENTAL BIOLOGY AND MEDICINE (2019) IF: 3.005
    Calcimimetic R568 improved cardiac remodeling by classic and novel renin-angiotensin system in spontaneously hypertensive rats:

    DOI: 10.1177/1535370219854325
    Sample: Plasm
  18. REPRODUCTIVE TOXICOLOGY (2021) IF: 3.143
    Morphine dependence and withdrawal-induced changes in mouse Sertoli cell (TM4) line: Evaluation of apoptotic, inflammatory and oxidative stress biomarkers

    DOI: 10.1016/j.reprotox.2021.09.004

    PMID: 34517100

    		 Sample: Tm4 Cells
  19. ACTA PHARMACOLOGICA SINICA (2017) IF: 3.223
    Osthole pretreatment alleviates TNBS-induced colitis in mice via both cAMP/PKA-dependent and independent pathways

    DOI: 10.1038/aps.2017.71
    Sample: Serum
  20. Annals of Translational Medicine (2020) IF: 3.297
    Teasaponin suppresses Candida albicans filamentation by reducing the level of intracellular cAMP

    DOI: 10.21037/atm.2020.01.124
    Sample: Yem30 Cell
  21. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY (2015) IF: 3.337
    Analysis and application of a neutralizing linear epitope on liable toxin B of enterotoxin Escherichia coli

    DOI: 10.1007/s00253-015-6448-x
    Sample: Epithelial Cell
  22. ACTA PHARMACOLOGICA SINICA (2018) IF: 3.562
    Osthole prevents tamoxifen-induced liver injury in mice

    DOI: 10.1038/s41401-018-0171-y

    PMID: 30315252

    		 Sample: Tissue Homogenate
  23. MOLECULAR AND CELLULAR ENDOCRINOLOGY (2018) IF: 3.563
    Curcumin restrains hepatic glucose production by blocking cAMP/PKA signaling and reducing acetyl CoA accumulation in high-fat diet (HFD)-fed mice

    DOI: 10.1016/j.mce.2018.02.018
    Sample: Liver,Primary Hepatocytes
  24. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS (2021) IF: 3.575
    Dopamine receptor D1 signaling stimulates lipolysis and browning of white adipocytes

    DOI: 10.1016/j.bbrc.2021.12.040

    PMID: 34953210

    		 Sample: Adipose Tissue,Adipocyte
  25. NEUROGASTROENTEROLOGY AND MOTILITY (2022) IF: 3.598
    Effect of phosphodiesterase-4 inhibitor rolipram on colonic hypermotility in water avoidance stress rat model

    DOI: 10.1111/nmo.14317

    PMID: 35037375

    		 Sample: Colon Tissue
  26. Frontiers in Aging Neuroscience (2019) IF: 3.633
    Royal Jelly Alleviates Cognitive Deficits and β-Amyloid Accumulation in APP/PS1 Mouse Model Via Activation of the cAMP/PKA/CREB/BDNF Pathway and Inhibition of Neuronal Apoptosis

    DOI: 10.3389/fnagi.2018.00428
    Sample: Tissue Homogenate
  27. JOURNAL OF NATURAL PRODUCTS (2020) IF: 3.782
    Eleutheroside E Enhances the Long-Term Memory of Radiation-Damaged C. elegans through G-Protein-Coupled Receptor and Neuropeptide Signaling Pathways

    DOI: 10.1021/acs.jnatprod.0c00650

    PMID: 33196193

    		 Sample: Tissue Homogenate
  28. TOXICOLOGY AND APPLIED PHARMACOLOGY (2017) IF: 3.791
    Biatriosporin D displays anti-virulence activity through decreasing the intracellular cAMP levels

    DOI: 10.1016/j.taap.2017.03.004
  29. EXPERIMENTAL CELL RESEARCH (2021) IF: 3.905
    Down-regulation of A3AR signaling by IL-6-induced GRK2 activation contributes to Th17 cell differentiation

    DOI: 10.1016/j.yexcr.2021.112482
    Sample: Cell Sample
  30. BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS (2022) IF: 4.117
    A2A R mediated modulation in IP3 levels altering the [Ca2+]i through cAMP-dependent PKA signalling pathway

    DOI: 10.1016/j.bbagen.2022.130242

    PMID: 36084905

    		 Sample: Hek 293 Cell
  31. ACS Chemical Neuroscience (2018) IF: 4.211
    The Inhibition by Guanfu Base A of Neuropathic Pain Mediated by P2Y12 Receptor in Dorsal Root Ganglia

    DOI: 10.1021/acschemneuro.8b00399

    PMID: 30475578

    		 Sample: Cell Culture Supernatant
  32. JOURNAL OF ETHNOPHARMACOLOGY (2021) IF: 4.36
    Protective effect of the combination of essential oil from patchouli and tangerine peel against gastric ulcer in rats

    DOI: 10.1016/j.jep.2021.114645

    PMID: 34530094

    		 Sample: Serum
  33. Scientific Reports (2021) IF: 4.38
    The protective effect of the PDE-4 inhibitor rolipram on intracerebral haemorrhage is associated with the cAMP/AMPK/SIRT1 pathway

    DOI: 10.1038/s41598-021-98743-w

    PMID: 34611179

    		 Sample: Brain Tissue
  34. Scientific Reports (2021) IF: 4.38
    Structure-based redesigning of pentoxifylline analogs against selective phosphodiesterases to modulate sperm functional competence for assisted reproductive technologies

    DOI: 10.1038/s41598-021-91636-y

    PMID: 34112881

    		 Sample: Sperm
  35. STEM CELLS AND DEVELOPMENT (2022) IF: 4.39
    cAMP agonist forskolin disrupts mitochondrial metabolism and induces senescence in human mesenchymal cells

    DOI: 10.1089/scd.2022.0180
    Sample: Mscs
  36. Frontiers in Neuroscience (2021) IF: 4.677
    Long Non-coding RNA Uc.48+ Small Interfering RNA Alleviates Neuroinflammatory Hyperalgesia in Gp120-Treated Rats via the P2Y12 Receptor

    DOI: 10.3389/fnins.2021.663962
    Sample: Hek 293 Cells
  37. MOLECULES (2022) IF: 4.927
    Effect of Natural Adenylcyclase/cAMP/CREB Signalling Activator Forskolin against Intra-Striatal 6-OHDA-Lesioned Parkinson’s Rats: Preventing Mitochondrial, Motor and Histopathological Defects

    DOI: 10.3390/molecules27227951
    Sample: Brain
  38. Metabolites (2022) IF: 4.932
    Serum Metabolomics Based on GC-MS Reveals the Antipyretic Mechanism of Ellagic Acid in a Rat Model

    DOI: 10.3390/metabo12060479

    PMID: 35736412

    		 Sample: Cerebrospinal Fluid (Csf)
  39. ENDOCRINOLOGY (2023) IF: 5.051
    Aspirin suppresses hepatic glucagon signaling through decreasing production of thromboxane A2

    DOI: 10.1210/endocr/bqac217
    Sample: Hepatocyte
  40. CNS Neuroscience & Therapeutics (2022) IF: 5.243
    Inhibition of phosphodiesterase-4 in the spinal dorsal horn ameliorates neuropathic pain via cAMP-cytokine-Cx43 signaling in mice

    DOI: 10.1111/cns.13807
    Sample: Spinal Cord
  41. Chinese Medicine (2021) IF: 5.455
    Anti-platelet aggregation of Panax notoginseng triol saponins by regulating GP1BA for ischemic stroke therapy

    DOI: 10.1186/s13020-021-00424-3

    PMID: 33468191

    		 Sample: Serum
  42. INTERNATIONAL JOURNAL OF ONCOLOGY (2019) IF: 5.65
    CD38 affects the biological behavior and energy metabolism of nasopharyngeal carcinoma cells

    DOI: 10.3892/ijo.2018.4651

    PMID: 30535454

    		 Sample: Cell Lysate
  43. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2022) IF: 6.208
    Conjunctival Fluid Secretion Impairment via CaCC-CFTR Dysfunction Is the Key Mechanism in Environmental Dry Eye

    DOI: 10.3390/ijms232214399
    Sample: Conjunctiva Epithelium
  44. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2022) IF: 6.208
    A Novel Furocoumarin Derivative, 5-((diethylamino)me-13 thyl)-3-phenyl-7H-furo [3,2-g] chromen-7-one Upregulates Melanin Synthesis via the Activation of cAMP/PKA and MAPKs Signal Pathway: In Vitro and In Vivo Study

    DOI: 10.3390/ijms232214190
    Sample: Skin
  45. Biomaterials Science (2019) IF: 6.843
    ONO-1301 loaded nanocomposite scaffolds modulate cAMP mediated signaling and induce new bone formation in critical sized bone defect

    DOI: 10.1039/C9BM01352K
    Sample: Mesenchymal Stem Cell
  46. PLOS BIOLOGY (2020) IF: 7.076
    Nucleotide de novo synthesis increases breast cancer stemness and metastasis via cGMP-PKG-MAPK signaling pathway

    DOI: 10.1371/journal.pbio.3000872

    PMID: 33186350

    		 Sample: Cell Sample
  47. EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY (2022) IF: 7.088
    Site-specific N-terminal PEGylation-based controlled release of biotherapeutics: An application for GLP-1 delivery to improve pharmacokinetics and prolong hypoglycemic effects

    DOI: 10.1016/j.ejmech.2022.114672

    PMID: 35973313

    		 Sample: Hek293 Cell
  48. BIOMEDICINE & PHARMACOTHERAPY (2022) IF: 7.419
    Phosphodiesterase 4B activation exacerbates pulmonary hypertension induced by intermittent hypoxia by regulating mitochondrial injury and cAMP/PKA/p-CREB/PGC-1α signaling

    DOI: 10.1016/j.biopha.2022.114095
    Sample: Serum,Pulmonary Arterial Smooth Muscle Cells (Pasm
  49. Cells (2022) IF: 7.666
    Forskolin, an Adenylcyclase/cAMP/CREB Signaling Activator Restoring Myelin-Associated Oligodendrocyte Destruction in Experimental Ethidium Bromide Model of Multiple Sclerosis

    DOI: 10.3390/cells11182771

    PMID: 36139346

    		 Sample: Brain Tissue
  50. Antioxidants (2022) IF: 7.675
    Muscone and (+)-Borneol Cooperatively Strengthen CREB Induction of Claudin 5 in IL-1β-Induced Endothelium Injury

    DOI: 10.3390/antiox11081455
    Sample: Bend.3 Cell
  51. Cell Death & Disease (2022) IF: 8.469
    Combining adoptive NK cell infusion with a dopamine-releasing peptide reduces senescent cells in aged mice

    DOI: 10.1038/s41419-022-04562-w

    PMID: 35383143

    		 Sample: Splenic Nk Cell
  52. Nature Communications (2022) IF: 17.694
    The gut microbe Bacteroides fragilis ameliorates renal fibrosis in mice

    DOI: 10.1038/s41467-022-33824-6

    PMID: 36241632

    		 Sample: Renal,Prtc Cells
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