ELISA Kit for Cyclic Adenosine Monophosphate (cAMP) Pan-species (General) Competition ELISA

c-AMP; 3'-5'-Cyclic Adenosine Monophosphate; Adenosine Cyclophosphate

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  • ELISA Kit for Cyclic Adenosine Monophosphate (cAMP) Packages (Simulation)
  • ELISA Kit for Cyclic Adenosine Monophosphate (cAMP) Packages (Simulation)
  • ELISA Kit for Cyclic Adenosine Monophosphate (cAMP) Results demonstration
  • CEA003Ge.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of Cyclic Adenosine Monophosphate (cAMP) and the recovery rates were calculated by comparing the measured value to the expected amount of Cyclic Adenosine Monophosphate (cAMP) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 84-94 88
EDTA plasma(n=5) 85-96 93
heparin plasma(n=5) 98-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cyclic Adenosine Monophosphate (cAMP) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Cyclic Adenosine Monophosphate (cAMP) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Cyclic Adenosine Monophosphate (cAMP) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 80-89% 94-103% 79-103% 89-101%
EDTA plasma(n=5) 92-105% 79-104% 89-97% 87-94%
heparin plasma(n=5) 88-102% 86-93% 80-102% 98-105%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1 Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
Reagent Diluent 1×300µL Stop Solution 1×6mL
TMB Substrate 1×9mL Instruction manual 1
Wash Buffer (30 × concentrate) 1×20mL

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 50µL standard or sample to each well.
    And then add 50µL prepared Detection Reagent A immediately.
    Shake and mix. Incubate 1 hour at 37°C;
3. Aspirate and wash 3 times;
4. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
5. Aspirate and wash 5 times;
6. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
7. Add 50µL Stop Solution. Read at 450 nm immediately.

ELISA Kit for Cyclic Adenosine Monophosphate (cAMP)

Test principle

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Cyclic Adenosine Monophosphate (cAMP) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Cyclic Adenosine Monophosphate (cAMP) and unlabeled Cyclic Adenosine Monophosphate (cAMP) (Standards or samples) with the pre-coated antibody specific to Cyclic Adenosine Monophosphate (cAMP). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Cyclic Adenosine Monophosphate (cAMP) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Cyclic Adenosine Monophosphate (cAMP) in the sample.

Citations

  • Cold exposure stimulates lipid metabolism, induces inflammatory response in the adipose tissue of mice and promotes the osteogenic differentiation of BMMSCs via the p38 MAPK pathwayPubMed: 26617802
  • Light Emitting Diodes Down‐reCavia (Guinea pig )lates Cathelicidin, Kallikrein, and Toll‐like Receptor 2 Expressions in Keratinocytes and Rosacea‐like Mouse SkinPubmed:27315464
  • P2Y12 Promotes Migration of Vascular Smooth Muscle Cells Through Cofilin Dephosphorylation During Atherogenesis.pubmed:28062501
  • Influences of Hyriopsis cumingii polysaccharides on mice immunosignaling molecules and T lymphocyte differentiation09540105.2017.1306494
  • Immunomodulatory effect of APS and PSP is mediated by Ca2+-cAMP and TLR4/NF-κB signaling pathway in macrophage.pubmed:27732877
  • Ginsenoside Rg5 Inhibits Succinate-Associated Lipolysis in Adipose Tissue and Prevents Muscle Insulin Resistance.pubmed:28261091
  • The relation of innate and adaptive immunity with viral-induced acute asthma attacks: Focusing on IP-10 and cathelicidinpubmed:27955890
  • Structural re-positioning, molecular modelling, oxidative degradation, and biological screening of linagliptin as adenosine 3 receptor (ADORA3) modulators …Pubmed:29768061
  • Astragaloside IV Inhibits Adipose Lipolysis and Reduces Hepatic Glucose Production via Akt Dependent PDE3B Expression in HFD-Fed MicePubmed:29410630
  • Dobutamine Up-Regulates Aquaporin 5 Expression in Septic Pulmonary Edema Model In Vitro via cAMP-PKA/CREB Signaling Pathway10.1166:nnl.2018.2602
  • Tuberculous meningitis is associated with higher cerebrospinal HIV-1 viral loads compared to other HIV-1-associated meningitidesPubmed:29394269
  • Developmental competence of buffalo (Bubalus bubalis) denuded oocytes cocultured with cumulus cells: Protective role of cumulus cellsPubmed:30092373
  • iTRAQ-Based Proteomics to Reveal the Mechanism of Hypothalamus in Kidney-Yin Deficiency Rats Induced by Levothyroxine
  • Calcimimetic R568 improved cardiac remodeling by classic and novel renin-angiotensin system in spontaneously hypertensive ratsPubmed: 31159562
  • Deletion of the serine protease CAP2/Tmprss4 leads to dysregulated renal water handling upon dietary potassium depletionPubmed: 31863073
  • Cytoplasm lipids can be modulated through hormone-sensitive lipase and are related to mitochondrial function in porcine IVM oocytesPubmed: 32172783
  • Astragaloside IV relieves gestational diabetes mellitus in genetic mice through reducing hepatic gluconeogenesisPubmed: 32160476
  • Altered absorptive function in the gall bladder during cholesterol gallstone formation is associated with abnormal NHE3 complex formationPubmed: 32557227
  • Antihypertensive effects of allicin on spontaneously hypertensive rats via vasorelaxation and hydrogen sulfide mechanismsPubmed: 32480217
  • DNA microarray analysis of hypothermia-exposed murine lungs for identification of forensic biomarkersPubmed: 33161360
  • Adrenomedullin 2 attenuates LPS-induced inflammation in microglia cells by receptor-mediated cAMP-PKA pathway33253929
  • A Green and Blue Monochromatic Light Combination Therapy Reduces Oxidative Stress and Enhances B-Lymphocyte Proliferation through Promoting ¡­33828639
  • Physiological crosstalk between the Mel1a and Mel1c pathways modulates melatonin-mediated, monochromatic light combination-induced bursa B-lymphocyte?¡­
  • The role of adrenergic and muscarinic receptors in stress-induced cardiac injury34245378
  • Sult2b1 deficiency exacerbates ischemic stroke by promoting pro-inflammatory macrophage polarization in mice34815805

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