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Competitive elisa

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The Competitive ELISA is a laboratory assay used to detect and quantify the presence of specific analytes in a sample. It operates on the principle of competitive binding, where the target analyte in the sample competes with a known quantity of labeled analyte for binding to a limited number of specific antibodies. The resulting signal intensity is inversely proportional to the concentration of the target analyte, allowing for its quantification.

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Lab products found in correlation

Tcs sp8 microscope, by Leica (1 mentions)

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Competitive ELISA kit Competitive ELISA assay Competitive cGMP ELISA kit Cyclic GMP competitive ELISA Kit ELISAs

7 protocols using competitive elisa

1

Plasma 8-Isoprostane ELISA Assay

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Plasma 8-isoprostane levels were measured as a marker of oxidative stress by competitive ELISA (Cayman Chemical, Ann Arbor).
Sinha R., Sinha I., Calcagnotto A., Trushin N., Haley J.S., Schell T.D, & Richie JP J.r. (2017). Oral supplementation with liposomal glutathione elevates body stores of glutathione and markers of immune function. European journal of clinical nutrition, 72(1), 105-111.
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2

Vibration-Induced Degranulation and Signaling

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Degranulation was determined as the percentage of β-hexosaminidase released into the media (Kuehn et al., 2010 ) 20 min after vibration. Alternatively, cells plated in DS-coated imaging chambers (Ibidi #80826), were vibrated for 5 min and anti-CD63-APC was added (1:20). Confocal images were acquired over time using a Leica TCS SP8 microscope and processed using ImageJ, selecting Region of Interest (ROI) Manager to determine anti-CD63-APC mean fluorescent intensity per cell.
PGD2 in cell-free supernatants or serum samples was determined by competitive ELISA (Cayman Chemicals). Phosphorylation of signaling proteins were determined by Western blotting after vibration for 5 min as described (Boyden et al., 2016 (link), Tkaczyk et al., 2002 (link)).
Naranjo A.N., Bandara G., Bai Y., Smelkinson M.G., Tobío A., Komarow H.D., Boyden S.E., Kastner D.L., Metcalfe D.D, & Olivera A. (2020). CRITICAL SIGNALING EVENTS IN THE MECHANOACTIVATION OF HUMAN MAST CELLS VIA P.C492Y-ADGRE2. The Journal of investigative dermatology, 140(11), 2210-2220.e5.
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3

Quantifying AII and ET-1 Levels

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The concentrations of angiotensin II (AII) and endothelin (ET)-1 were assayed in cell
cultures treated with UA using a commercially available competitive ELISA (Cayman
Chemical, USA). All assays were performed according to the manufacturer's protocols.
The absorbance of each sample was determined using an Ultra Microplate (Biotek, USA)
and reported as ng/mL and pg/mL.
Albertoni G, & Schor N. (2014). Resveratrol inhibits the intracellular calcium increase and angiotensin/endothelin system activation induced by soluble uric acid in mesangial cells. Brazilian Journal of Medical and Biological Research, 48(1), 51-56.
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4

Quantifying Oxidized Guanine Species

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DNA and RNA are damaged by oxidation, with guanine as most prone to oxidation. Using a competitive ELISA (Cayman, Ann Harbor, MI, USA), different oxidized guanine species were measured in urine including 8-hydroxyguanosine, 8-hydroxy-2'-deoxyguanosine, and 8-hydroxyguanine. We used these oxidized guanine species as marker for oxidized DNA and RNA. Oxidized DNA/RNA levels are reported in pg/ml.
Hagenbeek F.A., Roetman P.J., Pool R., Kluft C., Harms A.C., van Dongen J., Colins O.F., Talens S., van Beijsterveldt C.E., Vandenbosch M.M., de Zeeuw E.L., Déjean S., Fanos V., Ehli E.A., Davies G.E., Hottenga J.J., Hankemeier T., Bartels M., Vermeiren R.R, & Boomsma D.I. (2020). Urinary Amine and Organic Acid Metabolites Evaluated as Markers for Childhood Aggression: The ACTION Biomarker Study. Frontiers in Psychiatry, 11, 165.
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5

Carvedilol Modulates UV-Induced PGE2

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JB6 P+ cells were cultured in 12-well plates and pre-treated with carvedilol (5 μM) or the vehicle for 2 h and were then exposed to UV. After further incubation with the drug for 24 h post-UV, the culture media were collected from the plates and PGE2 levels determined by competitive ELISA, according to the manufacturer’s protocols (Cayman Chemical Co., Ann Arbor, MI, USA).
Chen M., Liang S., Shahid A., Andresen B.T, & Huang Y. (2020). The β-Blocker Carvedilol Prevented Ultraviolet-Mediated Damage of Murine Epidermal Cells and 3D Human Reconstructed Skin. International Journal of Molecular Sciences, 21(3), 798.
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6

UV-Induced PGE2 Measurement in JB6 Cells

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PGE2 activity was determined by culturing JB6 P+ cells in 12-well plates. The cells were pre-treated with or without carvedilol, S-, or R-carvedilol for 2 hours and were then exposed to 60 mJ/cm2 UV. After 24 hours of incubation, the culture media was removed from the plates, and PGE2 concentrations were determined via competitive ELISA according to the manufacturer’s protocol (Cayman Chemical Co., Ann Arbor, MI).
Liang S., Shamim M.A., Shahid A., Chen M., Cleveland K.H., Parsa C., Orlando R., Andresen B.T, & Huang Y. (2021). Prevention of skin carcinogenesis by the non-β-blocking R-carvedilol enantiomer. Cancer prevention research (Philadelphia, Pa.), 14(5), 527-540.
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7

Purification and Quantification of PGE2

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PGE2 was first extracted from pleural lavage by C18 column purification according to manufacturer’s directions (Cayman Chemical, Ann Arbor Michigan). PGE2 levels were determined by competitive ELISA (Cayman) according to manufacturer’s instructions.
Tucker T.A., Jeffers A., Boren J., Quaid B., Owens S., Koenig K.B., Tsukasaki Y., Florova G., Komissarov A.A., Ikebe M, & Idell S. (2016). Organizing empyema induced in mice by Streptococcus pneumoniae: effects of plasminogen activator inhibitor-1 deficiency. Clinical and Translational Medicine, 5, 17.
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