Multiscreenhts plates

Manufactured by Merck Group

Sourced in United States

MultiScreenHTS plates are a type of laboratory equipment designed for high-throughput screening (HTS) applications. They provide a platform for conducting multiple simultaneous assays in a compact, multi-well format. The core function of these plates is to facilitate efficient and organized experimentation, enabling researchers to screen large numbers of samples or compounds in a parallel manner.

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Lab products found in correlation

Fluoxetine, by Santa Cruz Biotechnology (2 mentions) Reserpine, by Merck Group (2 mentions) Tetrabenazine, by Merck Group (2 mentions) Vacuum manifold, by Merck Group (2 mentions) Unlabeled 5 ht, by Merck Group (2 mentions) Ne or trp, by PerkinElmer (2 mentions) Filter count scintillation fluid, by PerkinElmer (2 mentions) Ls6500, by Beckman Coulter (2 mentions) Red blood cell lysis buffer, by Merck Group (1 mentions) 40 m cell strainer, by BD (1 mentions) Ctl immunospot reader, by Cellular Technology (1 mentions) Concanavalin a (cona), by Thermo Fisher Scientific (1 mentions) Akp streptavidin, by BD (1 mentions) Penicillin, by Merck Group (1 mentions) Streptomycin, by Merck Group (1 mentions) Penicillin, by BD (1 mentions) Streptomycin, by BD (1 mentions) Alkaline phosphatase conjugated streptavidin, by Merck Group (1 mentions) Ks elispot, by Zeiss (1 mentions) Ionomycin, by Merck Group (1 mentions)

Close spelling variants of this product

MultiScreenHTS-HA 96-well filter plate (8 citations) MultiScreenHTS-HA filter plates (4 citations)

6 protocols using multiscreenhts plates

Microbial Uptake of Neurotransmitters

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T. sanguinis or B. theta was cultured anaerobically as described above and subcultured for 15–17 hours to reach stationary phase. Cells were washed and resuspended in Kreb-Ringer’s buffer, then left untreated or pre-treated with vehicle, unlabeled 5-HT (Sigma Aldrich), fluoxetine (Santa Cruz), reserpine (Sigma Aldrich) or tetrabenazine (Sigma Aldrich) for 30 min at 37°C. Cells were then incubated with 1 uM tritiated 5-HT (plus 0–500 uM unlabeled 5-HT for dose-dependent assays), NE or Trp (Perkin Elmer). For T. sanguinis, all uptake reactions were performed at room temperature. For B. theta, uptake reactions were performed at either room temperature or 37°C.
Uptake reactions were terminated by washing in ice cold Kreb-Ringer’s buffer. Washed T. sanguinis was trapped through 0.45 um PVDF filters on MultiScreenHTS plates connected to a vacuum manifold (EMD Millipore). For B. theta, washed cells were either trapped through 0.45 um PVDF filters or lysed by incubating in TE buffer plus triton and lysozyme. Extracted filters or lysed cells were incubated in 4 ml Filter Count scintillation fluid (Perkin Elmer) for 1 hour at room temp and radioactivity was measured using a liquid scintillation counter (Beckman LS6500).

Fung T.C., Vuong H.E., Luna C.D., Pronovost G.N., Aleksandrova A.A., Riley N.G., Vavilina A., McGinn J., Rendon T., Forrest L.R, & Hsiao E.Y. (2019). Intestinal serotonin and fluoxetine exposure modulate bacterial colonization in the gut. Nature microbiology, 4(12), 2064-2073.

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Microbial Uptake of Neurotransmitters

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Quantification of IFNγ-Producing Splenocytes

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The spleen was removed aseptically, pressed through a 40-µm cell strainer (Falcon), treated with Red Blood Cell lysis buffer (Sigma-Aldrich) to lyse erythrocytes, and washed with RPMI Medium 1640 supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 0.1 mg/ml streptomycin (all Thermo Fisher Scientific). Ethanol-activated MultiScreenHTS plates (Merck) were coated with purified rat anti-mouse IFNɣ antibody (clone R4-6A2, BD Bioscience) in PBS and blocked with RPMI Medium 1640 containing 10% FBS, 100 U/ml penicillin, and 0.1 mg/ml streptomycin. Splenocytes were seeded at 1 × 10⁶ cells/well and stimulated with 0.1 µg/ml peptide. Concanavalin A (Thermo Fisher Scientific) at the same concentration was used as an assay high control for IFNɣ production. After 20 h of incubation, IFNɣ-producing cells were detected with a biotinylated rat anti-mouse IFNɣ antibody (clone XMG1.2, BD Bioscience), AKP Streptavidin (BD Bioscience), and BCTP/NBT as a substrate (Thermo Fisher Scientific) for color development. The CTL ImmunoSpot Reader (Cellular Technology Ltd) was used for quantification of spots.

Ballhausen A., Przybilla M.J., Jendrusch M., Haupt S., Pfaffendorf E., Seidler F., Witt J., Hernandez Sanchez A., Urban K., Draxlbauer M., Krausert S., Ahadova A., Kalteis M.S., Pfuderer P.L., Heid D., Stichel D., Gebert J., Bonsack M., Schott S., Bläker H., Seppälä T., Mecklin J.P., Ten Broeke S., Nielsen M., Heuveline V., Krzykalla J., Benner A., Riemer A.B., von Knebel Doeberitz M, & Kloor M. (2020). The shared frameshift mutation landscape of microsatellite-unstable cancers suggests immunoediting during tumor evolution. Nature Communications, 11, 4740.

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Fluorocholic Acid Transport Inhibition

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Transport of 3β-fluorocholic acid (FCA) by the bile acid efflux pumps BSEP and MRP2 was determined by a competition assay with [³H]TC and [³H]EbG in BSEP and MRP2 membrane vesicles (Pharmtox). The assay was performed in 96-well plates. Each well contained 3.75 μg membrane vesicle, 10 mM MgCl₂, 4 mM ATP, 10 mM Tris, 250 mM sucrose, 0–10 mM FCA and 2 μM [³H]TC or [³H]EbG. The final volume in each well was 30 μL. The plates were incubated for 10 minutes at 37°C and uptake was halted by the addition of 150 μL ice-cold buffer solution. The contents of each well were transferred to a glass fiber filter plate (Multiscreen HTS plates; Merck Millipore) and washed 3 times with 200 μL of ice-cold buffer solution. Vesicles were lysed by incubation with 100 μL 0.1 M NaOH for 10 minutes at room temperature. An 80 μL aliquot of this solution was used for liquid scintillation counting. Data points were collected in triplicate. IC50-values were calculated with Graphpad Prism v3.00.

De Lombaerde S., Neyt S., Kersemans K., Verhoeven J., Devisscher L., Van Vlierberghe H., Vanhove C, & De Vos F. (2017). Synthesis, in vitro and in vivo evaluation of 3β-[18F]fluorocholic acid for the detection of drug-induced cholestasis in mice. PLoS ONE, 12(3), e0173529.

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B Cell Activation and Enumeration Assay

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Purified MZ B cells and FO B cells were cultured at 10⁵ cells/200 µl complete RPMI medium as described in the Cell sorting, cell culture, and adoptive transfer section. After 48 h of culture, cells were washed and distributed at various concentrations into Multiscreen HTS plates (EMD Millipore) precoated with 10 µg/ml goat anti–mouse IgM (H + L; Southern Biotech) in 2% FBS RPMI 1640 medium, and then incubated for 24 h at 37°C in 5% CO₂. Plates were washed, and then incubated with biotin-conjugated goat anti–mouse IgM (H + L; 1:5,000; Southern Biotechnology Associates) for 2 h at room temperature. Plates were washed, and then treated with alkaline phosphatase–conjugated Streptavidin (1:1,000; Sigma-Aldrich) and developed with 5-bromo-4-chloro-3-indoyl phosphate/NBT (Sigma-Aldrich). ISCs were enumerated using the KS ELISPOT (ZEISS).

Chen T.T., Tsai M.H., Kung J.T., Lin K.I., Decker T, & Lee C.K. (2016). STAT1 regulates marginal zone B cell differentiation in response to inflammation and infection with blood-borne bacteria. The Journal of Experimental Medicine, 213(13), 3025-3039.

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Murine IFN-γ ELISpot for Peptide Stimulation

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FA was always assessed 7 days after the last boost to compare immunizations. Splenocytes were plated in a range of concentrations (2 × 10³ to 1 × 10⁵/well) on Multiscreen HTS plates (Merck, NJ, USA) using the mouse IFN‐γ ELISpot BASIC (ALP) kit to detect murine IFN‐γ (Mabtech, Stockholm, Sweden). Cells were stimulated with titrated soluble peptide. As a positive control, cells were stimulated with PMA 50 ng/mL and Ionomycin 500 ng/mL (both Sigma‐Aldrich, St. Louis, MI). Background from control wells (unstimulated cells) were subtracted from the spot forming units counts. Counts were normalized to graph as a percentage of IFN‐γ producing cells and nonlinear regression curves were used to calculate EC_50.

Gilfillan C.B., Wang C., Mohsen M.O., Rufer N., Hebeisen M., Allard M., Verdeil G., Irvine D.J., Bachmann M.F, & Speiser D.E. (2019). Murine CD8 T‐cell functional avidity is stable in vivo but not in vitro: Independence from homologous prime/boost time interval and antigen density. European Journal of Immunology, 50(4), 505-514.

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