Direct Detection of Isolevuglandins in Tissues Using a D11 scFv-Alkaline Phosphatase Fusion Protein and Immunofluorescence

1.9K Views

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06:33 min

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July 5th, 2021

DOI :

10.3791/62603-v

July 5th, 2021

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Cassandra Warden*¹, Alan J. Simmons*², Lejla Pasic³, Ashley Pitzer⁴^,⁶, Sean S. Davies⁴, Justin H. Layer⁵, Raymond L. Mernaugh³, Annet Kirabo⁴^,⁶

¹Vanderbilt Eye Institute, Vanderbilt University Medical Center, ²Department of Cell and Developmental Biology, Vanderbilt University, ³Department of Biochemistry, Vanderbilt University, ⁴Division of Clinical Pharmacology, Department of Medicine, Vanderbilt University Medical Center, ⁵Division of Hematology and Oncology, Indiana University School of Medicine, ⁶Department of Molecular Physiology and Biophysics, Vanderbilt University

Summary

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Direct Detection

Isolevuglandins

D11 ScFv Alkaline Phosphatase

Immunofluorescence

Cardiovascular Disease

Tissue Detection Protocol

Antigen Retrieval

Paraffin Embedded Tissues

TBST Washing

Blocking Buffer

Alkaline Phosphatase Developer

Immunohistochemistry

Confocal Microscope

HEX Nuclear Stain

Chapters in this video

0:05

Introduction

0:54

Immunofluorescence

3:21

Negative Controls and Competitive Assays

4:37

Results: Detection of IsoLGs in Hypertensive and Normotensive Mice and Human Tissues

6:08

Conclusion

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