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Immunology and Infection
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监测人类痰样中的中性粒细胞和导管素 G 活性
JoVE Journal
Immunology and Infection
This content is Free Access.
JoVE Journal
Immunology and Infection
Monitoring Neutrophil Elastase and Cathepsin G Activity in Human Sputum Samples
Please note that all translations are automatically generated.
Click here for the English version.
监测人类痰样中的中性粒细胞和导管素 G 活性
DOI:
10.3791/62193-v
•
09:23 min
•
May 21, 2021
•
Dario L. Frey*
1,2
,
Matteo Guerra*
1,2,3,4
,
Marcus A. Mall
2,5,6,7
,
Carsten Schultz
3,8
1
Translational Lung Research Center Heidelberg (TLRC)
,
German Center for Lung Research (DZL)
,
2
Dept. of Translational Pulmonology
,
University of Heidelberg
,
3
Molecular Medicine Partnership Unit (MMPU), European Molecular Biology Laboratory (EMBL)
,
University of Heidelberg
,
4
Faculty of Biosciences, Collaboration for Joint Ph.D. Degree between EMBL and Heidelberg University
,
University of Heidelberg
,
5
Dept. of Pediatric Pulmonology, Immunology and Critical Care Medicine
,
Charité – Universitätsmedizin Berlin
,
6
Berlin Institute of Health (BIH)
,
7
German Center for Lung Research (DZL), Associated Partner Site, Berlin
,
8
Dept. of Chemical Physiology and Biochemistry
,
Oregon Health and Science University
Chapters
00:05
Introduction
01:07
Cell Isolation and Supernatant Separation
02:36
Quantification of Soluble Neutrophil Serine Protease (NSP) Activity via Fluorimeter or Plate Reader Assay
04:13
Membrane-bound NSPs Activity Measurement via Fluorescence Microscopy
05:30
Membrane-bound NSPs Activity Measurement via Flow Cytometry
07:25
Results: Representative Images and Quantification of Membrane-Bound NE Activity on Neutrophils Isolated from CF Patient Sputum
08:26
Conclusion
Summary
Automatic Translation
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Automatic Translation
本文所述的协议提供了一个指南,可视化和量化人类痰中中性粒细胞蛋白酶的活动。这种分析的应用范围从抗炎治疗的评估,到生物标志物验证、药物筛选和大队列临床研究。
Tags
Neutrophil Elastase
Cathepsin G
FRET Probe
Sputum
Airway Diseases
Cystic Fibrosis
Bronchiectasis
COPD
Plate Reader
Confocal Microscopy
Flow Cytometry
Salbutamol
Sputolysin
PBS
Cell Strainer
Centrifugation
Enzyme Standard Curve
FRET Probe NEmo-1
FRET Probe SSam
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