Multiplex Assay Kit for Cyclic Adenosine Monophosphate (cAMP) ,etc. by FLIA (Flow Luminescence Immunoassay) Pan-species (General) Multiplex ELISA

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

(Note: Up to 8-plex in one testing reaction)

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  • Multiplex Assay Kit for Cyclic Adenosine Monophosphate (cAMP) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Cyclic Adenosine Monophosphate (cAMP) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Cyclic Adenosine Monophosphate (cAMP) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA003Ge.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 84-93 87
EDTA plasma(n=5) 85-103 95
heparin plasma(n=5) 90-98 94
sodium citrate plasma(n=5) 85-104 92

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Cyclic Adenosine Monophosphate (cAMP) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 78-102% 97-105% 78-97% 95-102%
EDTA plasma(n=5) 83-97% 98-105% 79-94% 78-97%
heparin plasma(n=5) 83-98% 86-94% 81-98% 78-93%
sodium citrate plasma(n=5) 78-88% 89-102% 92-105% 79-99%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:cAMP) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 50μL standard or sample to each well,
    add 10μL magnetic beads,and 50μL Detection Reagent A,incubate 60min at 37°C on shaker;
3. Wash plate on magnetic frame for three times;
4. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
5. Wash plate on magnetic frame for three times;
6. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

Multiplex Assay Kit for Cyclic Adenosine Monophosphate (cAMP) ,etc. by FLIA (Flow Luminescence Immunoassay)

Test principle

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards,Labeled antigen and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest.A competitive inhibition reaction is launched between biotin labeled analytes of interest and unlabeled analytes of interest (Standards or samples) with the pre-coated antibody specific to analytes of interest. Following a wash to remove any unbound substances, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE) is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer. The MFI developed is reverse proportional to the concentration of analytes of interest in the sample.

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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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