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Oxiselect protein carbonyl elisa

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The OxiSelect™ Protein Carbonyl ELISA is a quantitative assay for measuring the levels of protein carbonyl content in biological samples. It provides a simple and efficient method for the detection and quantification of protein carbonyl formation, which is an indicator of oxidative stress.

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Oxiselect superoxide dismutase activity assay, by Cell Biolabs (1 mentions)

5 protocols using oxiselect protein carbonyl elisa

1

Quantifying Oxidative Stress Biomarkers

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Before the assays, placenta and fetal liver were homogenized in 1 ml of 0.15 M NaCl solution and centrifuged at 3000 rpm for 10 min at 4°C. Then, the supernatant was used for the analysis. The MDA concentrations in the serum, placenta, and fetus liver were quantified using the the thiobarbituric acid method [30] (link), which is based on the reaction of MDA with thiobarbituric acid to form a pink chromogen. Protein carbonyl levels were determined from reactions with dinitrophenylhydrazine, using a commercially available kit (Oxiselect protein carbonyl ELISA; Cell Biolabs, San Diego, CA, USA), according to the manufacturer's instructions. Absorbance was measured at 370 nm, and concentrations were calculated using a molar extinction coefficient of 22,000 M/cm at 370 nm after the subtraction of the blank absorbance. Data were normalized to protein content and expressed as nmol/mg protein.
Lin Y., Fang Z.F., Che L.Q., Xu S.Y., Wu D., Wu C.M, & Wu X.Q. (2014). Use of Sodium Butyrate as an Alternative to Dietary Fiber: Effects on the Embryonic Development and Anti-Oxidative Capacity of Rats. PLoS ONE, 9(5), e97838.
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2

Serum Protein Carbonylation in EAE

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Blood was isolated from EAE mice via terminal cardiac puncture, and serum levels of protein carbonylation were measured with OxiSelect™ Protein Carbonyl ELISA (Cell Biolabs) according to the manufacturer’s protocol. The absorbance at 450 nm was measured using a SpectraMax M5 microplate reader as described above.
Mendiola A.S., Ryu J.K., Bardehle S., Meyer-Franke A., Ang K.K., Wilson C., Baeten K.M., Hanspers K., Merlini M., Thomas S., Petersen M.A., Williams A., Thomas R., Rafalski V.A., Meza-Acevedo R., Tognatta R., Yan Z., Pfaff S.J., Machado M.R., Bedard C., Coronado P.E., Jiang X., Wang J., Pleiss M.A., Green A.J., Zamvil S.S., Pico A.R., Bruneau B.G., Arkin M.R, & Akassoglou K. (2020). Transcriptional profiling and therapeutic targeting of oxidative stress in neuroinflammation. Nature immunology, 21(5), 513-524.
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3

Protein Carbonyl ELISA for ROS Damage

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Carbonyl derivatives, a marker of ROS-mediated damage, were quantified by ELISA (OxiSelect Protein Carbonyl ELISA; Cell Biolabs; San Diego, CA). Mucus samples were analyzed based on equal protein loading (10 μg/mL) of the samples according to the manufacturer's instructions. ROS mediated damage was expressed as protein carbonyl nmol/mg.
Fishman J.E., Levy G., Alli V., Zheng X., Mole D.J, & Deitch E.A. (2014). The intestinal mucus layer is a critical component of the gut barrier that is damaged during acute pancreatitis. Shock (Augusta, Ga.), 42(3), 264-270.
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4

Measurement of Serum Biomarkers

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Blood samples were drawn into tubes with no additives, serum was aliquoted, immediately frozen, and stored at −80°C until analysis. The following kits were used: 25-hydroxyvitamin D ELISA (enzyme-linked immunosorbent assay) (Immunodiagnostics Systems, Scottsdale, AZ, USA), 1,25(OH)2 D ELISA (Immunodiagnostics Systems), testosterone ELISA (ALPCO Immunoassays, Salem, NH, USA), Total Antioxidant Capacity Assay, OxiSelect Superoxide Dismutase Activity Assay, OxiSelect MDA Adduct ELISA, OxiSelect Protein Carbonyl ELISA (Cell Biolabs, Inc., San Diego, CA, USA). The kits for measuring vitamin D metabolites employ highly specific 25(OH)D and 1,25(OH)2D3 antibodies. Solid-phase monoclonal anti-1,25(OH)2D3 was used for immunoextraction of 1,25(OH)2D3 prior to detection of the hormone by ELISA.
Scholten S.D., Sergeev I.N., Song Q, & Birger C.B. (2015). Effects of vitamin D and quercetin, alone and in combination, on cardiorespiratory fitness and muscle function in physically active male adults. Open Access Journal of Sports Medicine, 6, 229-239.
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5

Serum Protein Carbonylation in EAE

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Blood was isolated from EAE mice via terminal cardiac puncture, and serum levels of protein carbonylation were measured with OxiSelect™ Protein Carbonyl ELISA (Cell Biolabs) according to the manufacturer’s protocol. The absorbance at 450 nm was measured using a SpectraMax M5 microplate reader as described above.
Mendiola A.S., Ryu J.K., Bardehle S., Meyer-Franke A., Ang K.K., Wilson C., Baeten K.M., Hanspers K., Merlini M., Thomas S., Petersen M.A., Williams A., Thomas R., Rafalski V.A., Meza-Acevedo R., Tognatta R., Yan Z., Pfaff S.J., Machado M.R., Bedard C., Coronado P.E., Jiang X., Wang J., Pleiss M.A., Green A.J., Zamvil S.S., Pico A.R., Bruneau B.G., Arkin M.R, & Akassoglou K. (2020). Transcriptional profiling and therapeutic targeting of oxidative stress in neuroinflammation. Nature immunology, 21(5), 513-524.
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