NETosis Assay Kit (ab235979)
Key features and details
- Assay type: Quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Suspension cells
Overview
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Product name
NETosis Assay Kit -
Detection method
Colorimetric -
Sample type
Suspension cells -
Assay type
Quantitative -
Range
0.56 mU/well - 36 mU/well -
Species reactivity
Reacts with: Human -
Product overview
NETosis Assay Kit (ab235979) provides a simple and fast method for studying the process of NETosis ex vivo.
In this kit, primary neutrophils are stimulated to release NETs with either PMA or a calcium ionophore (both included). Unbound neutrophil elastase is washed away following NET generation. Following digest of NET DNA by S7 nuclease, the supernatant containing neutrophil elastase is added to a substrate, which is selectively cleaved by elastase to yield a 4-nitroaniline product that adsorbs light at 405 nm. Enough reagents are provided to test 24 sample conditions for NET production, with analysis in duplicate.
Note: This kit does not depend upon the DNA component of neutrophil extracellular traps (NETs), as DNA release can occur independently of NETosis.
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Platform
Microplate reader
Properties
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Storage instructions
Please refer to protocols. -
Components 1 kit 96-Well Plate (Colorimetric Assay) 1 unit 96-Well Plate Cover 1 unit A-23187 (25 mM) Assay Reagent 1 x 50µl Bovine Serum Albumin Assay Reagent 1 x 5g Calcium Chloride (1 M) Assay Reagent 1 x 1ml Cell-Based PMA (1 mM) 1 x 50µl EDTA (500 mM) Assay Reagent 1 x 1ml Human Neutrophil Elastase Assay Reagent 1 x 50µl NET Assay Neutrophil Elastase Substrate 1 x 250µl S7 Nuclease Assay Reagent 1 x 50µl
Images
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Example standard curve
Human neutrophil elastase standard curve.
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Measurement of released neutrophil elastase parallels measurement of released dsDNA.Human neutrophils were treated with PMA for 4 hours, washed and treated with S7 Nuclease for 15 minutes. The supernatant from each well was sampled and assayed for neutrophil elastase according to the protocol booklet. The supernatant was also tested for the prescence of soluble dsDNA using a green fluorescence probe.