Nanohead

Manufactured by PerkinElmer

The NanoHead is a high-precision laboratory instrument designed for advanced nanomaterial characterization. It utilizes state-of-the-art technology to provide accurate and reproducible measurements of particle size, shape, and distribution within nanoscale materials. The core function of the NanoHead is to enable researchers and scientists to gain a comprehensive understanding of the physical properties of nanoscale samples.

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Lab products found in correlation

Janus workstation, by PerkinElmer (1 mentions) Streptavidin coated donor beads, by PerkinElmer (1 mentions) Anti dykddddk mouse mab, by Fujifilm (1 mentions) Protein a coated acceptor beads, by PerkinElmer (1 mentions) Flexdrop precision reagent dispenser, by PerkinElmer (1 mentions) Envision plate reader, by PerkinElmer (1 mentions) Anti dykddddk tag monoclonal antibody, by Fujifilm (1 mentions) Viaflo, by Integra LifeSciences (1 mentions) Envision multilabel reader, by PerkinElmer (1 mentions) Cell explorer, by PerkinElmer (1 mentions) Celltiter glo reagent, by Promega (1 mentions)

3 protocols using nanohead

Thalidomide Binding Assay Using AlphaScreen

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We added 20 µl of bait mixture, containing 50 µM thalidomide and 0.5 µl of biotinylated HsCRBN in AlphaScreen buffer, to 384‐well AlphaPlates using a FlexDrop Precision Reagent Dispenser (PerkinElmer). We next added 0.8 µl of FLAG‐GST‐transcription factor proteins to 384‐well AlphaPlates using a NanoHead (PerkinElmer) and a Janus Workstation (PerkinElmer). After the 384‐well AlphaPlates were incubated at 26°C for 1 h, 5 µl of detection mixture containing 0.2 µg/ml anti‐DYKDDDDK mouse mAb (FUJIFILM Wako Pure Chemical), 0.08 µl of streptavidin‐coated donor beads and 0.08 µl of protein A‐coated acceptor beads (PerkinElmer) in AlphaScreen buffer were added to each well using a FlexDrop Precision Reagent Dispenser. After incubation at 26°C for 1 h, luminescent signals were detected using an EnVision plate reader (PerkinElmer).

Yamanaka S., Murai H., Saito D., Abe G., Tokunaga E., Iwasaki T., Takahashi H., Takeda H., Suzuki T., Shibata N., Tamura K, & Sawasaki T. (2021). Thalidomide and its metabolite 5‐hydroxythalidomide induce teratogenicity via the cereblon neosubstrate PLZF. The EMBO Journal, 40(4), e105375.

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High-Throughput Protein Interaction Screening

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All AlphaScreen reactions were conducted in AlphaPlate 384 titer plates (PerkinElmer). 20 ml of solution containing 0.5 μl of a biotin-tagged bait protein and 19.5 μl of the AlphaScreen buffer was dispensed into reaction plate by using automated multichannel pipet Viaflo and Viaflo Assist system (Integra). Next, 1 μl of each FLAG-GST tagged protein array protein was transferred from 384-well stock plate to the reaction plate using Janus automated dispensing workstation (PerkinElmer) and Nanohead, a 384-well micro syringe head (PerkinElmer). Then 9 μl of detection mixture containing 0.02 μl of anti-DYKDDDDK tag monoclonal antibody (Wako), 0.06 μl of streptavidin-conjugated AlphaScreen donor beads and 0.06 μl of protein A–conjugated AlphaScreen acceptor beads (PerkinElmer) in the AlphaScreen buffer were added to the reaction plates using FlexDrop dropper. After incubation at 25°C for 1 h in a dark incubator, AlphaScreen signal was detected by Envision multilabel reader (PerkinElmer).

Nishiyama K., Maekawa M., Nakagita T., Nakayama J., Kiyoi T., Chosei M., Murakami A., Kamei Y., Takeda H., Takada Y, & Higashiyama S. (2021). CNKSR1 serves as a scaffold to activate an EGFR phosphatase via exclusive interaction with RhoB-GTP. Life Science Alliance, 4(9), e202101095.

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High-throughput drug screening in cells

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High throughput drug screening was performed by the high throughput screening facility at the Columbia Genome Center. Briefly, we seeded cells into 384 well microplates at a concentration of 4000 cells per well in 50 μL of RPMI media supplemented with 10% FBS. The next day, we delivered compounds using an automated liquid handling station (Cell::Explorer, Perkin Elmer). We transferred compounds in triplicate (1 μM, DMSO) to assay plates using nano-head (Perkin Elmer). After 72 hours, we performed a cell viability assay using Cell Titer Glo reagent (Promega #G7573).
For data normalization, we used Pipeline Pilot Lab Analytics 17.1.0 and Chemistry 17.1.0 Collections. We used the Percent of Positive Control (PPC) normalization component where each well is expressed as a percentage of the average of the DMSO-only wells in the same plate.

Rashkovan M., Albero R., Gianni F., Perez-Duran P., Miller H.I., Mackey A.L., Paietta E.M., Tallman M.S., Rowe J.M., Litzow M.R., Wiernik P.H., Luger S., Sulis M.L., Soni R.K, & Ferrando A.A. (2022). Intracellular cholesterol pools regulate oncogenic signaling and epigenetic circuitries in Early T-cell Precursor Acute Lymphoblastic Leukemia. Cancer discovery, 12(3), 856-871.

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