Fluorescence-Based Ratiometric Measurement of CRAC Channel Activity in STIM-Orai-Overexpressing HEK-293 Cells. Chapter

abstract

• Calcium influx through store-operated Ca2+ entry (SOCE), mediated by STIM-operated Orai channels, is crucial for many cellular functions. To dissect the molecular mechanisms underlying the process of STIM-Orai activation and identify regulators that modify this process, ratiometric imaging of SOCE responses in HEK cells overexpressing STIM and Orai is a routinely used method. Here we describe one commonly used procedure of monitoring SOCE activity with a ratiometric membrane-permeable dye fura-2-AM. Other ratiometric indicators suitable for SOCE measurements are also discussed.

authors

• Zhou, Yubin

author list (cited authors)

• Zhang, S., He, L., Zhou, Y., & Wang, Y.

citation count

• 1

complete list of authors

• Zhang, Shuce||He, Lian||Zhou, Yubin||Wang, Youjun

Book Title

• The CRAC Channel

publication date

• September 2018

publisher

• Springer New York  Publisher

published in

• Methods Mol Biol  Journal

keywords

• Calcium
• Calcium Release Activated Calcium Channels
• Calcium Release-activated Calcium (crac) Channel
• Calcium Signaling
• Data Analysis
• FURA-2
• Fluorescent Dyes
• Fura-4f
• Fura-6f
• Fura-red
• Gem-geco1
• Gene Expression
• HEK293 Cells
• Humans
• Image Processing, Computer-Assisted
• Indo-1
• Microscopy, Fluorescence
• Molecular Imaging
• Ratiometric Ca2+ Measurements
• Software
• Store-operated Ca2+ Entry (soce)
• Stromal Interaction Molecules

PubMed Central ID

• 30203274

Digital Object Identifier (DOI)

• 10.1007/978-1-4939-8704-7_2

International Standard Book Number (ISBN) 13

• 9781493987023

start page

• 17

end page

• 39

volume

• 1843