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cGMP(Cyclic GMP) ELISA Kit

  • Cat.No.:E-EL-0083

  • Reactivity: Universal

To Purchase E-EL-0083

Size:
  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
Price: $495
Qty:

Product Details

Properties

Assay type Competitive-ELISA
Format 96T/48T
Assay time 2.0h
Detection range 0.78-50 pmol/mL
Sensitivity 0.47 pmol/mL
Sample type &Sample volume serum, plasma and other biological fluids; 50μL
Specificity This kit recognizes cGMP in samples. No significant cross-reactivity or interference between cGMP 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 cGMP 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 cGMP. During the reaction, cGMP in the sample or standard competes with a fixed amount of cGMP on the solid phase supporter for sites on the Biotinylated Detection Ab specific to cGMP. 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 cGMP 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℃, 6 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), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°C, 6 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 cGMP were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level cGMP 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 (pmol/mL) 98.40 299.90 999.00 92.40 326.30 1055.70
Standard deviation 5.20 13.50 35.00 4.80 18.60 53.80
CV (%) 5.28 4.50 3.50 5.19 5.70 5.10

Recovery

The recovery of cGMP 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) 91-104 96
EDTA plasma(n=8) 91-108 99
Cell culture media(n=8) 86-97 91

Target Information

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. Frontiers in Immunology (2022) IF: 8.786
    Sildenafil, a Type-5 Phosphodiesterase Inhibitor, Fails to Reverse Myeloid-Derived Suppressor Cell-Mediated T Cell Suppression in Cells Isolated From Tuberculosis Patients

    DOI: 10.3389/fimmu.2022.883886

    PMID: 35935981

    Sample: MDSC
  2. 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
  3. ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY (2022) IF: 7.129
    Effect of bisphenol F on sexual performance and quality of offspring in Male Wistar rats

    DOI: 10.1016/j.ecoenv.2022.114079

    PMID: 36115153

    Sample: Penile
  4. BIOMEDICINE & PHARMACOTHERAPY (2022) IF: 6.53
    Glutamine restores testicular glutathione-dependent antioxidant defense and upregulates NO/cGMP signaling in sleep deprivation-induced reproductive dysfunction in rats

    DOI: 10.1016/j.biopha.2022.112765

    PMID: 35247715

    Sample: Penile
  5. 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
  6. Frontiers in Endocrinology (2021) IF: 5.555
    Inhibition of Phosphodiesterase 5 Promotes the Aromatase-Mediated Estrogen Biosynthesis in Osteoblastic Cells by Activation of cGMP/PKG/SHP2 Pathway

    DOI: 10.3389/fendo.2021.636784

    Sample: UMR106 cell
  7. 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)
  8. ACS Omega (2022) IF: 4.132
    Ajuga bracteosa Exerts Antihypertensive Activity in l-NAME-Induced Hypertension Possibly through Modulation of Oxidative Stress, Proinflammatory Cytokines, and the Nitric Oxide/Cyclic Guanosine Monophosphate Pathway

    DOI: 10.1021/acsomega.2c03888

    PMID: 36157749

    Sample: serum
  9. AMERICAN JOURNAL OF PHYSIOLOGY-ENDOCRINOLOGY AND METABOLISM (2021) IF: 4.31
    Bee bread mitigates downregulation of steroidogenic genes, decreased spermatogenesis, and epididymal oxidative stress in male rats fed with high-fat diet

    DOI: 10.1152/ajpendo.00093.2021

    PMID: 34229480

    Sample: serum,testis
  10. JOURNAL OF ETHNOPHARMACOLOGY (2021) IF: 4.36
    Ziziphus Oxyphylla hydro-methanolic extract ameliorates hypertension in L-NAME induced hypertensive rats through NO/cGMP pathway and suppression of oxidative stress related inflammatory biomarkers.

    DOI: 10.1016/j.jep.2021.114825

    Sample: Serum
  11. 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
  12. REPRODUCTION (2020) IF: 3.206
    Orlistat reverses intratesticular lactate transport decline and infertility in male obese rats

    DOI: 10.1530/REP-20-0381

    PMID: 33112813

    Sample: testis
  13. ACTA PHARMACOLOGICA SINICA (2017) IF: 3.223
    Osthole prevents acetaminophen-induced liver injury in mice

    DOI: 10.1038/aps.2017.129

    Sample: Tissue homogenate
  14. BEHAVIOURAL BRAIN RESEARCH (2022) IF: 3.352
    Prenatal chronic stress impairs the learning and memory ability via inhibition of the NO/cGMP/PKG pathway in the Hippocampus of offspring

    DOI: 10.1016/j.bbr.2022.114009

    PMID: 35850398

    Sample: hippocampal tissue
  15. 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
  16. PLoS One (2022) IF: 3.752
    L-Arginine reverses maternal and pre-pubertal codeine exposure-induced sexual dysfunction via upregulation of androgen receptor gene and NO/cGMP signaling

    DOI: 10.1371/journal.pone.0274411

    PMID: 36099318

    Sample: penile
  17. American Journal of Translational Research (2022) IF: 3.94
    Dexlansoprazole prevents pulmonary artery hypertension by inhibiting pulmonary artery smooth muscle cell to fibroblast transition.

    PMID: 36105026

    Sample: plasma
  18. BMC Cardiovascular Disorders (2020) IF: 2.078
    Aggravated endothelial endocrine dysfunction and intimal thickening of renal artery in high-fat diet-induced obese pigs following renal denervation

    DOI: 10.1186/s12872-020-01472-7

    PMID: 32295540

    Sample: Tissue homogenate
  19. Biomed Research International (2018) IF: 2.583
    C-Type Natriuretic Peptide Ameliorates Lipopolysaccharide-Induced Cardiac Dysfunction in Rats with Pulmonary Arterial Hypertension

    DOI: 10.1155/2018/2813025

    Sample: plasma
  20. Pharmacological Reports (2019) IF: 2.761
    Simultaneous inhibition of neprilysin and activation of ACE2 prevented diabetic cardiomyopathy

    DOI: 10.1016/j.pharep.2019.05.008

    Sample: heart tissue,plasma
  21. CURRENT ISSUES IN MOLECULAR BIOLOGY (2023) IF: 2.976
    Effect of C-Type Natriuretic Peptide (CNP) on Spermatozoa Maturation in Adult Rat Epididymis

    DOI: 10.3390/cimb45020108

    Sample: epididymal tissue
  22. Molecular & Cellular Toxicology (2022) IF: 1.08
    FXYD3 promotes the proliferation, migration, and invasion of pancreatic cancer cells by regulating the cGMP-PKG signaling pathway

    DOI: 10.1007/s13273-021-00207-9

    Sample: pancreatic tissue,BxPC-3 cell,CAPAN2 cell
  23. 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
  24. Toxicology Reports (2021)
    Sildenafil augments fetal weight and placental adiponectin in gestational testosterone-induced glucose intolerant rats

    DOI: 10.1016/j.toxrep.2021.06.011

    Sample: Placenta
  25. Heliyon (2021)
    Sildenafil ameliorates leptin resistance and normalizes lipid handling in the hypothalamic and adipose tissues of testosterone-exposed pregnant rats

    DOI: 10.1016/j.heliyon.2021.e07574

    Sample: plasma
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