Monitoring Neutrophil Elastase and Cathepsin G Activity in Human Sputum Samples

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15.5K Views

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09:23 min

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May 21st, 2021

DOI :

10.3791/62193-v

May 21st, 2021

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Dario L. Frey*¹^,², Matteo Guerra*¹^,²^,³^,⁴, Marcus A. Mall¹^,²^,⁵^,⁶^,⁷, Carsten Schultz¹^,³^,⁸

¹Translational Lung Research Center Heidelberg (TLRC), German Center for Lung Research (DZL), ²Dept. of Translational Pulmonology, University of Heidelberg, ³Molecular Medicine Partnership Unit (MMPU), European Molecular Biology Laboratory (EMBL), University of Heidelberg, ⁴Faculty of Biosciences, Collaboration for Joint Ph.D. Degree between EMBL and Heidelberg University, University of Heidelberg, ⁵Dept. of Pediatric Pulmonology, Immunology and Critical Care Medicine, Charité – Universitätsmedizin Berlin, ⁶Berlin Institute of Health (BIH), ⁷German Center for Lung Research (DZL), Associated Partner Site, Berlin, ⁸Dept. of Chemical Physiology and Biochemistry, Oregon Health and Science University

Podsumowanie

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Enzyme Standard Curve

Rozdziały w tym wideo

0:05

Introduction

1:07

Cell Isolation and Supernatant Separation

2:36

Quantification of Soluble Neutrophil Serine Protease (NSP) Activity via Fluorimeter or Plate Reader Assay

4:13

Membrane-bound NSPs Activity Measurement via Fluorescence Microscopy

5:30

Membrane-bound NSPs Activity Measurement via Flow Cytometry

7:25

Results: Representative Images and Quantification of Membrane-Bound NE Activity on Neutrophils Isolated from CF Patient Sputum

8:26

Conclusion

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