JoVE Journal

Genetics

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Nanyang Technological University

Using a GFP-tagged TMEM184A Construct for Confirmation of Heparin Receptor Identity

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8.0K Views

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10:41 min

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February 17th, 2017

DOI :

10.3791/55053-v

February 17th, 2017

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Sara Lynn N. Farwell¹, Joshua B. Slee², Yaqiu Li¹, Linda J. Lowe-Krentz¹

¹Department of Biological Sciences, Lehigh University, ²Department of Natural Science, DeSales University

Podsumowanie

A construct encoding TMEM184A with a GFP tag at the carboxy-terminus designed for eukaryotic expression, was employed in assays designed to confirm the identification of TMEM184A as a heparin receptor in vascular cells.

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TMEM184A Construct

Heparin Receptor

Functional Identification

Rhodamine Heparin

Confocal Microscopy

Fluorescence Imaging

Rozdziały w tym wideo

0:05

Title

0:57

Vascular Cell GFP-TMEM184A Expression and Rhodamine-Heparin Binding

3:42

Fluorescence Resonance Energy Transfer (FRET) from GFP-TMEM184A to Rhodamine-Heparin, and Live Cell Imaging of Rhodamine-Heparin Uptake

5:15

In Vitro Heparin Binding Assay

8:07

Results: Representative GFP-TMEM184A-transfection Effects

9:44

Conclusion

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