Brand New Configuration for Your FRET and Fluorescent Protein Assays
- Built to meet your needs
- 3 lasers (V:405/B:488/Y:561 nm), 20 colors (V7/B6/Y7)
- Available with or without a small particle detector
- Perfect for fluorescent proteins, FRET, and marine biology assays
- Same all-around capabilities: integrated sample loader (deep-well plates compatible), up to 100,000 events/sec acquisition rate, high-throughput mode, superior sensitivity and stability, and seamless integration with agnostic API
ZE5 Cell Analyzer Violet-Blue-Yellow-Green (VBY) Configuration and Reagent* Guide
*Not a comprehensive list of fluorophores and fluorescent proteins that can be used with the ZE5 Cell Analyzer
|460/22||SBV440, Pacific Blue, mTurquoise|
|525/50||SBV515, BV510, AmCyan, CFP, Cascade Yellow|
|509/24||FITC, BB515, eGFP, mClover|
|692/80||SBB700, PerCp-Cy5.5, BB700, PE-Cy5|
|589/15||dsRed, tdTomato, RFP|
|615/24||PE-Dazzle, PE-CF594, mCherry|
FRET (Fluorescence Resonance Energy Transfer) has been broadly used to analyze protein interaction and protein assembly in living cells. The technique is based upon the transfer of energy from an excited donor fluorophore to an acceptor fluorophore, provided that: 1) the donor and acceptor are in close proximity (<10 nm) and 2) the emission spectra of the donor overlaps with the excitation of the acceptor. Traditionally, FRET assays are done by fluorescence microscopy, which is technically challenging, tedious, nonquantitative, and does not allow analysis in high-throughput or in large cell numbers.
In recent years, flow cytometry has been established as a more suitable technology for FRET assays, due to its high speed and high-throughput capabilities which overcome these limitations. The continuous advancement in FRET technologies and FRET pairs has further broadened flow cytometry-based FRET applications, especially in cancer research and high-throughput screening arenas.
“Using the imaging flow cytometer as described here, it takes 8 hr to analyze a 96-well plate with approximately 20,000 gated cells per well. However, we also routinely perform DAmFRET with standard (nonimaging) cytometers (ZE5 Cell Analyzer) that achieve much higher throughput (up to 20 samples per min).”
See how high-throughput DAmFRET assay was used to assess self-assembly of human inflammasome proteins, and how the ZE5 Cell Analyzer has taken screening throughput to the next level.
Take a Look at Some of the ZE5 FRET Publications
Polymerization of the caspase recruitment domain (CARD)-containing proteins in immune cells:
Holliday MJ et al. (2019). Structures of autoinhibited and polymerized forms of CARD9 reveal mechanisms of CARD9 and CARD11 activation. Nat Commun 10, 3070.
Cell death induced by toxic protein aggregates can be neutralized by co-assembly with antidote proteins:
Nuckolls N et al. (2020). The wtf4 meiotic driver utilizes controlled protein aggregation to generate selective cell death. Elife 9:e55694.
Assessing self-assembly of human inflammasome proteins with high-throughput DAmFRET assays:
Venkatesan S et al. (2019). Detecting and characterizing protein self-assembly in vivo by flow cytometry. J Vis Exp. (149).
Commonly Used FRET Pairs and Fluorescent Proteins
To help you get started, here are some commonly used FRET pairs and fluorescent proteins that you could use on the VBY model of the ZE5 Cell Analyzer.
|Donor/Acceptor||ZE5 Excitation Laser||ZE5 FRET Detector|
|eCFP/eYFP||405 nm||405 nm-525/50|
|mTurquoise-mCitrine||405 nm||405 nm-525/50|
|mTurquoise-mVenus||405 nm||405 nm-525/50|
|mAmetrine-tdTomato||405 nm||405 nm-615/24|
|mClover-mRuby||488 nm||488 nm-583/30|
|eGFP-mCherry||488 nm||488 nm-615/24|
|CyOFP1-mCardinal||488 nm||488 nm-692/80|
Bajar BT et al. (2016). A guide to fluorescent protein FRET pairs. Sensors (Basel). Publications16(9):1488.
|405 nm (V)||488 nm (B)||561 nm (Y)|
|eCFP||eGFP||dsRed, tdTomato, RFP|
Bajar BTet al. (2016). A guide to fluorescent protein FRET pairs. Sensors (Basel). publications16(9):1488.
Discover the ZE5 Cell Analyzer
With a dozen configuration and upgrade options, the ZE5 Cell Analyzer delivers the flexibility and efficiency that you need.Learn more about ZE5 applications ▸ View ZE5 Cell Analyzer product page ▸
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