Ab119553

Manufactured by Abcam

Sourced in United Kingdom

Ab119553 is a rabbit monoclonal antibody that recognizes the human GAPDH protein. It is intended for use in various immunochemical techniques.

Automatically generated - may contain errors

Lab products found in correlation

7600 chemistry analyzer, by Hitachi (1 mentions) Ab193695, by Abcam (1 mentions) Spectramax id3, by Molecular Devices (1 mentions) Cell death elisa kit, by Roche (1 mentions) Mithras multimode reader, by Berthold Technologies (1 mentions) Bms2038inst, by Thermo Fisher Scientific (1 mentions) Ab267628, by Abcam (1 mentions) Modulus luminometer, by Promega (1 mentions) Micro reduced glutathione gsh assay kit, by Solarbio (1 mentions) Ab119605, by Abcam (1 mentions) Ab108882, by Abcam (1 mentions)

5 protocols using ab119553

Urine Biomarkers in Nephrotic Syndrome

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In SSNS patients, two urine samples were collected during the nephrotic and remission phases. In the healthy controls, one urine sample was obtained from each individual; 24 h urine samples were collected and stored at −80°C until analysis. Urinary proteins were measured by the pyrogallol end-point method (10 (link)), while urinary GAGs excretion was examined using the modified Whiteman process (10 (link),11 (link)). GAGs levels were calculated using a calibration curve with HS as a standard, and corrected with urinary creatinine (Cr).
At the same time, blood samples (4 ml) were collected from SSNS patients and healthy controls and stored in tubes containing EDTA to prevent coagulation, and subsequently centrifuged at 400 × g for 5 min at 4°C. Enzymatic activity of Ela was quantified by ELISA according to the manufacturer's protocols (ab119553; Abcam, London, Cambridge, UK).
Urine levels of Creatinine (Cr) were measured using an Hitachi 7600 Chemistry Analyzer (Hitachi, Ltd., Tokyo, Japan).

Zhai S., Hu L., Zhong L., Tao Y, & Wang Z. (2017). Low molecular weight heparin may benefit nephrotic remission in steroid-sensitive nephrotic syndrome via inhibiting elastase. Molecular Medicine Reports, 16(6), 8613-8618.

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Quantifying Complement and Leukocyte Activation

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To determine complement and leukocyte activation, an enzyme-linked immunosorbent assay (ELISA) was performed. The SF, Hep/SF films, and PTFE sheets were pre-incubated with PBS at 37 °C for 5 min. The PBS was replaced with lithium heparinized blood and incubated at 37 °C for 1 h. Blood performed accordingly without samples was used as a control. After incubation, the plasma was isolated by centrifugation at 1400× g for 15 min. Complement activation was determined by using ELISA Kits for complement fragments C3a (ab279352, Abcam, Cambridge, UK) and C5a (ab193695, Abcam, Cambridge, UK). Leukocyte activation was determined by using ELISA Kits for polymorphonuclear neutrophil (PMN) elastase (ab119553, Abcam, Cambridge, UK). Measurements were performed according to the manufacturer’s instructions and in quadruplicate for each sample. The absorbance was measured using a microplate reader (SpectraMax® iD3, Molecular Devices, LLC., San Jose, CA, USA) [39 (link),40 (link)].

Fungmongkonsatean T., Jongjitwimol J., Paensuwan P., Nuamchit T., Siriwittayawan D., Kanokpanont S., Damrongsakkul S, & Thitiwuthikiat P. (2022). Hemocompatibility Evaluation of Thai Bombyx mori Silk Fibroin and Its Improvement with Low Molecular Weight Heparin Immobilization. Polymers, 14(14), 2943.

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Quantifying Neutrophil Degranulation Markers and NETs

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Dedicated enzyme-linked immunosorbent assay (ELISA) kits were used for quantitative measurement of neutrophil degranulation markers, including polymorphonuclear neutrophil (PMN)-elastase (ab119553; Abcam, Cambrige, UK), neutrophil gelatinase-associated lipocalin (NGAL) (KIT 036RUO; BioPorto Diagnostics A/S, Hellerup, Denmark), and MPO (BMS2038INST; Invitrogen, Waltham, MA, US).
NETs-associated MPO–DNA complexes were measured by adding serum into 96-well plates coated with a monoclonal anti-human MPO antibody (MABX4043-10KC; Millipore, Burlington, MA, US) after saturation of no specific binding site with bovine serum albumin, followed by incubation with peroxidase-labelled anti-DNA monoclonal antibody included in the Cell Death ELISA kit (11774425001; Roche Diagnostics, Mannheim, Germany). The optical absorbance was measured at 405 wavelength by using Berthold Mithras multimode reader (Berthold Technologies GmbH, Bad Wildbad, Germany).

Buso G., Faggin E., Bressan A., Galliazzo S., Cinetto F., Felice C., Fusaro M., Erdmann A., Pauletto P., Rattazzi M, & Mazzolai L. (2023). Biomarkers of Neutrophil Activation in Patients with Symptomatic Chronic Peripheral Artery Disease Predict Worse Cardiovascular Outcome. Biomedicines, 11(3), 866.

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Quantifying Plasma Biomarkers of Neutrophil Extracellular Traps

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We collected 5ml of peripheral blood from individuals in cohort 3, The cells are removed by centrifugation 3000g for 5min. The supernatant, designated plasma is carefully collected from the cell pellet using a Pasteur pipette. Levels of calprotectin (S100A8/A9) and glutathione in plasma samples were analyzed using a commercial enzyme-linked immunosorbent assay (ELISA) (ab267628, Abcam, USA) kit and Micro Reduced Glutathione (GSH) Assay Kit (BC1175, Solarbio, Beijing) respectively. Then elastase and cell-free DNA (cfDNA) were included for detecting NET (23 (link)). The level of Elastase was quantified via ELISA (ab119553, Abcam, USA). For the detection of cell-free DNA (cfDNA), we used PicoGreen dsDNA Assays Kits (P7589, Thermo Fisher Scientific, China) to quantify the cfDNA, the protocol is as follows: 1. Dilute the concentrated PicoGreen^® Dye stock two-hundred (200) fold with 1×TE; 2. Prepare Standard Curve with 1mg Deoxyribonucleic acid from calf thymus Type XV(D4522-1MG, Sigma, USA); 3. Measurement of fluorescence via Modulus Luminometer (9200-003, Turner BioSystems) and calculate cfDNA concentrations.

Cai M.L., Gui L., Huang H., Zhang Y.K., Zhang L., Chen Z, & Sheng Y.J. (2022). Proteomic Analyses Reveal Higher Levels of Neutrophil Activation in Men Than in Women With Systemic Lupus Erythematosus. Frontiers in Immunology, 13, 911997.

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Quantifying Immune Markers in COVID-19

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Enzyme-linked immunosorbent assay (ELISA) was performed according to manufacturer’s protocol to quantify myeloperoxidase (MPO) (Abcam, ab119605), lactoferrin (Abcam, ab108882) and elastase levels (Abcam, ab119553) in plasma from healthy donors, non-COVID-19 ARDS, COVID-19 patients and cell media supernatant of resiquimod treated NDN cultures from healthy donors (15 μM, Sigma-Aldrich). Briefly, a standard curve with known concentrations of the corresponding enzymes was generated to obtain their equivalent optical densities in a colorimetric assay. The concentration of the relevant enzymes in each sample was inferred from their optical densities by using the standard curve.

Reyes L., A. Sanchez-Garcia M., Morrison T., Howden A.J., Watts E.R., Arienti S., Sadiku P., Coelho P., Mirchandani A.S., Zhang A., Hope D., Clark S.K., Singleton J., Johnston S., Grecian R., Poon A., McNamara S., Harper I., Fourman M.H., Brenes A.J., Pathak S., Lloyd A., Blanco G.R., von Kriegsheim A., Ghesquiere B., Vermaelen W., Cologna C.T., Dhaliwal K., Hirani N., Dockrell D.H., Whyte M.K., Griffith D., Cantrell D.A, & Walmsley S.R. (2021). A type I IFN, prothrombotic hyperinflammatory neutrophil signature is distinct for COVID-19 ARDS. Wellcome Open Research, 6, 38.

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