384 well glass bottom microplate

Manufactured by Greiner

Sourced in Germany

The 384-well glass-bottom microplate is a laboratory equipment designed for various experimental applications. It features a grid of 384 individual wells with a glass bottom, providing optical transparency for imaging and analysis purposes.

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Lab products found in correlation

Lsm 880, by Zeiss (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions)

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384-well microplate (25 citations) Glass bottoms (3 citations)

2 protocols using 384 well glass bottom microplate

Quantifying Protein-Protein Interactions

Cited 1 time

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Five microlitres of glutathione Sepharose 4B beads slurry (GE Healthcare, Buckinghamshire, UK) were mixed with 20 µg of GST-fused bait proteins (GST-Cx43^WT_NT or GST-Cx43^W4A+L7A_NT) and incubated on a rotating wheel at 4 °C for at least 30 min. Subsequently the beads were washed twice with 150 mM NaCl, 50 mM Tris at pH 7.4 and resuspended in 6 µL of the same buffer. Of a 5 µM prey solution (GFP-LC3B or GFP-GABARAP), 10 µL was plated into the well of a 384-well glass-bottom microplate (Greiner Bio-One, Frickenhausen, Germany), after which 10% of the previous beads solution was added. Samples were then imaged on a confocal microscope. To quantify the protein recruitment to beads the maximum brightness along a straight line drawn through a single bead was taken (maximal fluorescence). Next, the average brightness of an empty portion of each picture was measured (background fluorescence) and subtracted from the maximal fluorescence for each bead [16 (link)].

Catarino S., Ribeiro-Rodrigues T.M., Sá Ferreira R., Ramalho J., Abert C., Martens S, & Girão H. (2020). A Conserved LIR Motif in Connexins Mediates Ubiquitin-Independent Binding to LC3/GABARAP Proteins. Cells, 9(4), 902.

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YTHDC1 Phase Separation Assay

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Before droplet formation experiments, purified MBP-YTHDC1-EGFP protein stock was incubated with TEV protease (lab stock at 200:1 molar ratio) for 2 hours on ice to cleave off the MBP tag. All phase separation assays were performed in the phase separation buffer (20 mM HEPES 7.3, 300 mM NaCl, 1mM MgCl₂, 1mM DTT, 5% Dextran T500 (Pharmacosmos).
The YTHDC-EGFP1 proteins were centrifuged at 13,000 g for 5 min to remove small protein pellets and diluted into the desired concentrations with phase separation buffer. The RNA samples were also diluted into the desired concentrations in phase separation buffer. To start liquid droplet formation, 5 μL YTHDC-EGFP1 protein sample was mixed with 5 μL RNA sample into a 384-well glass-bottom microplate (Greiner bio-one) pre-coated with 1 mg/ml BSA (Sigma). The final concentration of proteins and RNAs are indicated in the figures. The droplets were visualized with confocal microscopy (ZEISS LSM 880). The time-lapse data were processed using FIJI/ImageJ.

Cheng Y., Xie W., Pickering B.F., Chu K.L., Savino A.M., Yang X., Luo H., Nguyen D.T., Mo S., Barin E., Velleca A., Rohwetter T., Patel D.J., Jaffrey S.R, & Kharas M.G. (2021). N6-methyladenosine on mRNA facilitates a phase-separated nuclear body that suppresses myeloid leukemic differentiation. Cancer cell, 39(7), 958-972.e8.

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