Bioraptr frd

Manufactured by Beckman Coulter

The BioRAPTR FRD is a laboratory instrument designed for automated sample preparation. It provides a platform for various sample processing tasks, including liquid handling, mixing, and incubation. The BioRAPTR FRD is a versatile tool that can be utilized in a range of laboratory applications.

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Lab products found in correlation

Viewlux plate reader, by PerkinElmer (1 mentions) Envision plate reader, by PerkinElmer (1 mentions) Digitonin, by Merck Group (1 mentions) Pintool, by Fujifilm (1 mentions) Celltiter glo ctg luminescent cell viability assay, by Promega (1 mentions) Multidrop combi, by Thermo Fisher Scientific (1 mentions) Envision multilabel plate reader, by PerkinElmer (1 mentions) Multidrop combi dispenser, by Thermo Fisher Scientific (1 mentions)

4 protocols using bioraptr frd

High-Throughput Screening of HDAC Inhibitors

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HCT116, HEK293, HepG2, and SU-DHL-6 cells were seeded into 1536-well white solid bottom assay plates at desired cell densities in 5 μL medium using a microfluidic reagent dispenser (BioRAPTR FRD, Beckman Coulter, Indianapolis, IN). Before plating 5 μL of human neural stem cells into each well, assay plates were pre-coated with 5 μL of 10 μg/mL fibronectin in culture medium at 37°C for one hour and the coating solution was removed by centrifuging the inverted plates. Cells were settled overnight in a cell incubator at 37°C, 5%CO₂, followed by addition of 23 nL of compound solution on a pintool workstation (Kalypsys, San Diego, CA). After compound incubation for indicated period at 37°C/5% CO₂, 5 μL of HDAC-Glo was dispensed using BioRAPTR FRD. To measure cell viability, 5 μL of CellTiter-Glo Cell Viability reagent was added to each well in a separate assay plate using BioRAPTR FRD. The luminescence signals of HDAC-Glo and CellTiter-Glo assays were measured on a ViewLux plate reader (Perkin-Elmer, Shelton, CT).

Hsu C.W., Shou D., Huang R., Khuc T., Dai S., Zheng W., Klumpp-Thomas C, & Xia M. (2016). Identification of HDAC inhibitors using a cell-based HDAC I/II assay. Journal of biomolecular screening, 21(6), 643-652.

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Acetylcholinesterase Inhibition Assay

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The AChE inhibition assay was described in our previous study.^{21 (link)} Briefly, recombinant human AChE (50 mU/mL) was dispensed (4 µL/well) into black clear-bottom 1536-well plates. Chlorpyrifos-oxon and BW284C51, known nonselective and selective AChE inhibitors, respectively, were used as positive controls. Twenty-three nanoliters of test compounds with concentrations ranging from 0.37 nM to 28.75 μM and controls were transferred into the assay plates using a Wako Pintool. After a 30 min incubation period at room temperature, 4 µL of colorimetric detection cocktail solution (DTNB, acetylthiocholine) was added to each well using a BioRaptr FRD (Beckman Coulter). The final DMSO concentration in the assay well was 0.29%. Assay plates were incubated at room temperature for another 10 min, followed by measuring the absorbance readout (405 nm) using an Envision plate reader (PerkinElmer).

Li S., Li A.J., Travers J., Xu T., Sakamuru S., Klumpp-Thomas C., Huang R, & Xia M. (2021). Identification of Compounds for Butyrylcholinesterase Inhibition. Slas Discovery, 26(10), 1355-1364.

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High-Throughput Screening of IFN-β Modulators

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300 HLA-B HiBit cells/well were plated into white 1536-well plates (Greiner, 789173-F) in 5 uL/well of GM using a Multidrop Combi (Thermo Scientific). Column 1 did not contain IFN-β as a control, and all other columns contained 4.0 ng/mL IFN-β, where column 4 was used as IFN-β only control. 23 nL/well DMSO was added to columns 1–4 while 23 nL/well compounds in DMSO were added to columns 5–48 using a pintool (Wako). For counter screening, digitonin (Sigma, D141) in DMSO was added to column 2 at a final concentration of 92 uM as a cytotoxic control. For Nano-Glo HiBit Lytic Detection System (HiBit assay) (Promega, N3040) and CellTiter-Glo Luminescent Cell Viability Assay (CTG) (Promega, G7572), 2.5 uL/well reagent was added with a BioRAPTR FRD (Beckman Coulter), plates were incubated in the dark at ambient temperature for 10 min, and luminescence measured with a ViewLux 1430 Ultra HTS (Perkin Elmer) running Wallac 1430 Manager and Explorer software v3.02. Two of the three primary screening libraries (NPC and MIPE5.0) were pinned, incubated, and assayed on our in-house Kalypsys robotics system. IFN-β titration in Figure 2 was dispensed at 50 nl/well with a Mosquito (TTP Labtech). Refer to supplemental materials and methods for detailed protocol tables as we have described previously.^{63 (link)}

Kinder T.B., Dranchak P.K, & Inglese J. (2020). High throughput screening to identify inhibitors of the type I interferon – major histocompatibility complex class I pathway in skeletal muscle. ACS chemical biology, 15(7), 1974-1986.

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High-throughput Akt phosphorylation assay

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Neuro 2A cells were seeded (4,000 cells/well) in 4 µL of DMEM with 0.5% FBS and 1x Penicillin/Streptomycin in white solid bottom tissue-culture treated 1536-well plates (Greiner, 789173-F) with MultiDrop Combi Dispenser (Thermo Scientific). The cells were incubated for 24 h at 37 °C, 5% CO₂, 95% humidity. Twenty-three nL of tested compounds were transferred to each well with an automated pin-tool station (Kalypsys). Following incubation for 15 min at 37 °C, 0.5 µL IGF (500 ng/mL of final concentration) was added and incubated for additional 40 min at 37 °C. After the addition of 1.5 µL 4x lysis buffer and 15-min incubation at room temperature (25 °C), 2 µL of HTRF^® conjugated antibodies (fluorophore d2 conjugated anti-Akt monoclonal antibody and Europium³⁺ cryptate-labeled anti-pS473-Akt monoclonal antibody, obtained from CisBio, 1:80 dilution) were added with BioRAPTR FRD™ (Beckman Coulter). The plates were incubated at room temperature for 20 h. The signal was measured by EnVision Multilabel Plate Reader (PerkinElmer) with excitation at 330 nm, emission at 620 nm (donor) and 665 nm (acceptor). Results were calculated and analyzed in a format of the ratio 665 nm/620 nm multiplied by 10⁴.
Relative activity was calculated by normalizing each HTRF signal from each sample well to the mean HTRF signal from the DMSO-only control wells.

Huang B.X., Newcomer K., Kevala K., Barnaeva E., Zheng W., Hu X., Patnaik S., Southall N., Marugan J., Ferrer M, & Kim H.Y. (2017). Identification of 4-phenylquinolin-2(1H)-one as a specific allosteric inhibitor of Akt. Scientific Reports, 7, 11673.

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