For the NaCl concentration series, in vitro reactions were prepared as described above in 50 mM Tris (pH 7.5), 1 mM DTT, and the indicated NaCl concentration (25, 100, 250, 500 mM, or 1 M). 12.5 μM PTBP3 was incubated with 0 ng/μL RNA (PTBP3 alone) or 15 ng/μL LE RNA (PTBP3 & vg1 LE). For the RNA concentration series, in vitro reactions were prepared as described above in 50 mM Tris (pH 7.5), 100 mM NaCl, 1 mM DTT. 12.5 μM PTBP3 (PTBP3 & LE) or buffer controls (LE alone,) were incubated with the indicated RNA concentration of LE RNA (0 ng/μL, 15 ng/μL, 37.5 ng/μL, and 75 ng/μL). Turbidity was assayed in a 384-well glass bottom plate (MatTek Corporation) with 20 μL samples sealed with clear optical film (Excel Scientific) to prevent evaporation. Absorbance of the samples at 600 nm was read using a Cytation 5 Multi-Mode Reader (BioTek) after 1 h of incubation at room temperature. Absorbance data was normalized to buffer and TEV protease controls. In the turbidity assays, no fluorescent labels were used for either the protein or RNA.
384 well glass bottom plate
Manufactured by MatTek
The 384-well glass bottom plate is a laboratory equipment designed for cell-based assays and high-throughput screening applications. It features a glass bottom that allows for high-quality imaging and improved imaging performance compared to standard plastic plates. The plate has 384 individual wells, providing a high-density format for efficient use of limited sample volumes and workspace. The glass bottom ensures optimal optical clarity and minimizes interference with microscopy techniques.
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Lab products found in correlation
2 protocols using 384 well glass bottom plate
1
PTBP3 Binding to vg1 LE RNA Assayed by Turbidity
2
Modulating Phase Separation of PTBP3
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For the NaCl concentration series, in vitro phase separation reactions were prepared as described above in 50 mM Tris (pH 7.5), 1 mM DTT, and the indicated NaCl concentration (25 mM, 100 mM, 250 mM, 500 mM, or 1 M). 12.5 µM PTBP3 was incubated with 0 ng/µL RNA (PTBP3 alone) or 15 ng/µL LE RNA (PTBP3 & vg1 LE). For the RNA concentration series, in vitro phase separation reactions were prepared as described above in 50 mM Tris (pH 7.5), 100 mM NaCl, 1 mM DTT. 12.5 µM PTBP3 (PTBP3 & LE) or buffer controls (LE alone,) were incubated with the indicated RNA concentration of LE RNA (0 ng/µL, 15 ng/µL, 37.5 ng/µL, and 75 ng/µL). Turbidity was assayed in a 384-well glass bottom plate (MatTek Corporation) with 20 µL samples sealed with clear optical film (Excel Scientific) to prevent evaporation. Absorbance of the samples at 600 nm was read using a Cytation 5 Multi-Mode Reader (BioTek) after 1 hour of incubation at room temperature. Absorbance data was normalized to buffer and TEV protease controls. In the turbidity assays, no fluorescent labels were used for either the protein or RNA.
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