Total RNA was extracted from cell cultures 4 days after induced differentiation in vitro (unless indicated elsewhere) using the RNeasy Plus Mini Kit (QIAGEN), and reverse transcribed using the iScript cDNA synthesis kit (Biorad) according to the manufacturer’s instructions. ELANE mRNA was quantified using real-time Q-PCR with GAPDH as an internal control. For quantification of ELANE protein level, whole cell lysates were collected after 10-day neutrophil maturation culture which were then subject to ELISA analysis using the PMN (neutrophil) elastase human ELISA kit (ThermoFisher) strictly following the manufacturer’s instructions. All gene expression data represent the mean of at least three biological replicates.
Pmn neutrophil elastase human elisa kit
Manufactured by Thermo Fisher Scientific
The PMN (neutrophil) elastase human ELISA kit is a laboratory tool used to measure the concentration of PMN elastase, an enzyme found in neutrophils, in human samples. The kit provides a quantitative assay for the detection and measurement of PMN elastase levels.
Automatically generated - may contain errors
2 protocols using pmn neutrophil elastase human elisa kit
1
Quantifying ELANE mRNA and Protein
2
Quantification of Plasma Biomarkers in Blood Samples
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Blood was processed within 60 minutes of withdrawal. Blood was centrifuged twice (1000 x g followed by 10000 x g, both for 10 min at 4°C) to obtain platelet-free plasma and stored in aliquots at -80°C. Plasma MPO (Human Myeloperoxidase Quantikine ELISA Kit, Bio-Techne) and DNA-histone complexes (Cell Death Detection ELISA, Sigma-Aldrich) were quantified according to the manufacturer’s recommendations. NE levels were assessed with the Human Sepsis Magnetic Bead Panel 3 (EMD Millipore) in a Luminex MagPix instrument (Thermo Fisher Scientific) for AAA study samples or with PMN (Neutrophil) Elastase Human ELISA Kit (Thermo Fisher Scientific) for endotoxemia plasma samples. Citrullinated histone H3 (citH3) was measured as reported elsewhere [32 (link)]. Differential blood count was determined with a Sysmex XN-350 device. Biochemical parameters were assessed by the Vienna General Hospital routine laboratory.
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