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JoVE Journal

Immunology and Infection

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A Simple and Efficient Method to Detect Nuclear Factor Activation in Human Neutrophils by Flow Cytometry

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08:26 min

April 9th, 2013

April 9th, 2013

17,720 Views

0:05

Title

1:25

Isolation of Neutrophils (PMN) from Human Blood

3:02

Activating PMN

4:03

Isolation and Fixation of Nuclei

4:51

Nuclei Immunolabeling for Flow Cytometry Analysis

6:15

Flow Cytometry Analysis

6:49

Results: Monitoring NF-κB Dynamics

7:54

Conclusion

Neutrophils are the most abundant leukocytes in blood. Neutrophils possess transcriptionally regulated functions such as production of proinflammatory cytokines and inhibition of apoptosis. These functions can be studied with the method presented here, which allows detection and quantification of nuclear factors by flow cytometry in isolated nuclei

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