Flx 800 fluorescence plate reader

Manufactured by Agilent Technologies

The FLx-800 is a fluorescence plate reader manufactured by Agilent Technologies. It is designed to measure fluorescent signals in multi-well microplates. The device utilizes excitation and emission wavelengths to detect and quantify fluorescent molecules in biological samples.

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Lab products found in correlation

Uric acid, by Merck Group (1 mentions) Sodium borohydride, by Merck Group (1 mentions) Oxalic acid, by Merck Group (1 mentions) Diethylenetriamine, by Merck Group (1 mentions) Oxaloacetic acid, by Merck Group (1 mentions) Terephthalaldehyde, by Merck Group (1 mentions) Quick rna miniprep kit, by Zymo Research (1 mentions) Glass bottom 96 well plate, by MatTek (1 mentions) Truseq stranded mrna sample preparation kit, by Illumina (1 mentions) 2200 tapestation system, by Agilent Technologies (1 mentions) Novaseq 6000 system, by Illumina (1 mentions) D1000 screentape, by Agilent Technologies (1 mentions) Qubit dsdna hs assay kit, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Fluorescence plate reader (76 citations)

3 protocols using flx 800 fluorescence plate reader

Fluorescent Copper Sensor Synthesis

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Diethylenetriamine,
terephthalaldehyde, copper(II) nitrate (pentahydrate), sodium borohydride,
and solvents were obtained from Sigma Aldrich. All acids used in the
specificity test were obtained from Fisher Scientific, with the exception
of oxalic acid, oxaloacetic acid, and uric acid, which were obtained
from Sigma Aldrich. Highest purity available was used in every case,
and HPLC-grade solvents were used. All black 96 well assay plates
(Microfluor 2Black Microtiter) were obtained from Thermo Scientific.
All plates were read with a Bio-Tek FLx-800 fluorescence plate reader
(BioTek, Winooski, VT) on the “top read” setting with
sensitivity set to 60. NMR was performed on a Bruker Avance III Ultrashield
400 Plus spectrometer (Bruker BioSpin Corporation, Billerica, MA).
Mass spectrometry was performed on an Advion expression CMS Electrospray
Ionization instrument (Advion, Ithica, NY). All graphs and associated
calculations were produced and performed using GraphPad Prism software
(San Diego, CA).

Hontz D., Hensley J., Hiryak K., Lee J., Luchetta J., Torsiello M., Venditto M., Lucent D., Terzaghi W., Mencer D., Bommareddy A, & VanWert A.L. (2020). A Copper(II) Macrocycle Complex for Sensing Biologically Relevant Organic Anions in a Competitive Fluorescence Assay: Oxalate Sensor or Urate Sensor?. ACS Omega, 5(31), 19469-19477.

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Quantifying RNA-binding Compound Levels

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DM1 myotubes were grown in Mat-Tek 96-well glass bottom plates, differentiated, and treated with 5 μM 2H-K2-Pro or 2H-K4NMeS as described above for 24 h. Cells were then washed with 100 μL of 1× DPBS and then lysed in 30 μL of RNA Lysis Buffer from a Zymo Quick RNA Miniprep Kit. Intrinsic fluorescence intensity of the RNA-binding modules was measured using a BioTek FLX-800 fluorescence plate reader (excitation wavelength: 360/340 nm; emission wavelength: 460/440 nm; sensitivity = 90), and concentrations of compound were extrapolated to standard curves of 2H-K2-Pro or 2H-K4NMeS spiked into untreated cell lysate. n = 6 replicates; 2 independent experiments.

Benhamou R.I., Abe M., Choudhary S., Meyer S.M., Angelbello A.J, & Disney M.D. (2020). Optimization of the linker domain in a dimeric compound that degrades an r(CUG) repeat expansion in cells. Journal of medicinal chemistry, 63(14), 7827-7839.

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Illumina-based Transcriptome Profiling Protocol

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NGS libraries were prepared with the Illumina TruSeq Stranded mRNA Sample Preparation kit (Illumina, #20020595) according to the manufacturer’s instructions (Illumina TruSeq Stranded mRNA Reference Guide, # 1000000040498 v00, October 2017), using 400 ng of total RNA as input and applying 12 cycles of PCR at the final enrichment step. The produced NGS libraries were quality controlled using the D1000 ScreenTape and D1000 reagents (Agilent, #5067-5582 and 5067-5583) on the Agilent 2200 TapeStation system. Quantification of the libraries was performed using the Qubit dsDNA HS Assay Kit (Thermo Fisher, #Q32854) on the Biotek FLx800 Fluorescence plate reader. Paired-End 51 bp data were produced using the Illumina NovaSeq 6000 system, generating 26 million reads per sample on average.
The sequencing data were base called with Illumina’s Real-Time Analysis (RTA v3.4.4) software and processed into FASTQ file format, containing the sequence data, with the bcl2fastq2-v2.20.0.422 tool.

Steinhauser S., Estoppey D., Buehler D.P., Xiong Y., Pizzato N., Rietsch A., Wu F., Leroy N., Wunderlin T., Claerr I., Tropberger P., Müller M., Davison L.M., Sheng Q., Bergling S., Wild S., Moulin P., Liang J., English W.J., Williams B., Knehr J., Altorfer M., Reyes A., Mickanin C., Hoepfner D., Nigsch F., Frederiksen M., Flynn C.R., Fodor B.D., Brown J.D, & Kolter C. (2024). The transcription factor ZNF469 regulates collagen production in liver fibrosis. bioRxiv.

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