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Preprint

CitH3‐DNA ELISA

YS Yuanlin Song
QZ Qiaoliang Zhu
JZ Jian Zhou

Last updated date: Nov 2, 2025 Views: 23 Forks: 0

original An abbreviated version of this protocol was published in Pulmonary Circulation
Lianhua Qingke ameliorates lipopolysaccharide‐induced lung injury by inhibiting neutrophil extracellular traps formation and pyroptosis
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  1. Coating: Microtiter 96-well plates were coated overnight at 4°C with 5 μg/mL of anti-citH3 (ab5103, Abcam) as the capture antibody.
  2. Washing: Plates were washed three times with washing buffer.
  3. Sample and Antibody Incubation: 20 μL of BALF samples and 80 μL of incubation buffer containing a peroxidase-labeled anti-DNA monoclonal antibody (Cell Death Detection ELISA Kit, Roche; 1:25 dilution) were added to each well.
  4. Incubation: The plate was incubated for 2 hours at room temperature with shaking at 300 rpm.
  5. Washing: Plates were washed three times with washing buffer.
  6. Substrate Addition: 100 μL of peroxidase substrate (ABTS) was added to each well.
  7. Detection: After incubating for 20 minutes at room temperature in the dark, the absorbance was measured at a wavelength of 405 nm.
Copyright: © 2025 The Author(s); This is an open-access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/).
How to cite：
Readers should cite both the Bio-protocol preprint and the original research article where this protocol was used:
  1. Song, Y, Zhu, Q and Zhou, J(2025). CitH3‐DNA ELISA. Bio-protocol Preprint. bio-protocol.org/prep2870.
  2. Peng, W., Qi, H., Zhu, W., Tong, L., Rouzi, A., Wu, Y., Han, L., He, L., Yan, Y., Pan, T., Liu, J., Wang, Q., Jia, Z., Song, Y., Zhu, Q. and Zhou, J.(2023). Lianhua Qingke ameliorates lipopolysaccharide‐induced lung injury by inhibiting neutrophil extracellular traps formation and pyroptosis. Pulmonary Circulation 13(4). DOI: 10.1002/pul2.12295

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