Microtest 96

Manufactured by BD

Sourced in United States

The Microtest 96 is a laboratory equipment designed for performing microplate-based assays. It provides a standardized platform for conducting various experiments and tests requiring a 96-well format. The Microtest 96 is a versatile tool that can be utilized across different research and testing applications.

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Lab products found in correlation

Celltiter 96 aqueous one solution cell proliferation assay, by Promega (12 mentions) Ovcar3, by American Type Culture Collection (10 mentions) Mda mb 435, by American Type Culture Collection (9 mentions) Mda mb 231, by American Type Culture Collection (7 mentions) Celltiter blue cell viability assay, by Promega (7 mentions) Mtt proliferation assay kit, by Cayman Chemical (2 mentions) Synergy mx, by Agilent Technologies (1 mentions) Celltiter blue assay kit, by Promega (1 mentions) Synergy 2 microplate reader, by Agilent Technologies (1 mentions) Spectramax m2 multi mode microplate reader, by Molecular Devices (1 mentions)

Close spelling variants of this product

Microtest 96-well plate Flat-black-bottom 96-well microtest plate Microtest 96 plates Microtest

28 protocols using microtest 96

Inhibition of Aflatoxin Production

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For the purification of substances that can inhibit the production of AFs, a small-scale assay system with a 96-well flat-bottom tissue culture plate (Microtest 96; Becton Dickinson, Franklin Lakes, NJ, USA) was used [14 (link)]. After the GY agar medium (100 μL) was solidified in each well of the culture plate, an aliquot (10–20 μL) of each fraction at each purification step of the inhibitory substance(s) was then applied onto the resulting GY agar in the well. Spore suspension of A. parasiticus NFRI-95 was inoculated onto the resulting agar medium and cultured at 28 °C for 2 or 3 days. The color of the fungal mycelia was observed from the underside of the plate. A decrease in the intensity of the NA red color or loss of the NA red color indicated that the tested fraction contained the inhibitory activity.

Ichinomiya M., Fukushima-Sakuno E., Kawamoto A., Nakagawa H., Hatabayashi H., Nakajima H, & Yabe K. (2022). Inhibition of Aflatoxin Production by Citrinin and Non-Enzymatic Formation of a Novel Citrinin-Kojic Acid Adduct. Journal of Fungi, 9(1), 29.

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Millimeter-Wave Exposure System Design

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The exposure system consisted of two main units: i) an exposure chamber, and ii) a signal generator unit (Fig. 1).
Exposure chamber. All exposures were carried out inside a MEMMERT UNE 400 incubator (Memmert, Schwabach, Germany) set at 32°C. Samples under test (SUTs), described in the following section, were placed into one well of a 12-well tissue culture plate (TCP) (353072, Microtest 96, Becton Dickinson, Franklin Lakes, NJ) made of polystyrene.
Each well was 22.09 mm in diameter. Exposures were performed from the bottom by an open-ended rectangular waveguide (WG) WR15 (aperture size 3.81 × 1.905 mm²) located 5 mm from the bottom of TCP. The latter was set on a 5 mm-thick plastic support with a 3.5 cm hole in diameter, centered with the exposed well. The open-ended WG was connected to an MMW generator (QuinStar Technology, Torrance, CA) by means of standard WR15 waveguides (their total length is 27.5 cm).

Orlacchio R., Zhadobov M., Alekseev S.I., Nikolayev D., Sauleau R., Le Page Y, & Le Dréan Y. (2019). Millimeter-Wave Heating in In Vitro Studies: Effect of Convection in Continuous and Pulse-Modulated Regimes. Bioelectromagnetics, 40(8).

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Cytotoxicity of Compounds against Cancer Cell Lines

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Human melanoma cancer cells (MDA-MB-435), human breast cancer cells (MDA-MB-231) and human ovarian cancer cells (OVCAR3) were obtained from the American Type Culture Collection. The cell lines were cultured in RPMI 1640 medium containing fetal 10% bovine serum, 100 units/mL penicillin and 100 μg/mL streptomycin. The cells were grown at 37 °C under 5% CO₂, and then harvested during the log-phase growth by trypsinization followed by two washes to remove all traces of enzymes. Cells were seeded in 96-well clear, flat-bottom plates (Microtest 96, Falcon) at a density of 5000 cells per well. Each plate was incubated overnight at 37 °C under 5% CO₂. Samples dissolved in DMSO were diluted and added to the appropriate wells to give final concentrations of 20, 4, 0.8, 0.16, and 0.032 μM for pure compounds, and a total volume of 100 μL and 0.5% DMSO per well. The cells with the test samples were then incubated for 72 h at 37 °C. Cell viability was examined using a commercial absorbance assay (CellTiter 96 AQ_ueous One Solution Cell Proliferation Assay, Promega Corp). IC₅₀ values were determined as the concentration required to diminish cellular growth by 50% compared to the untreated controls after 72 h of continuous exposure. Taxol (paclitaxel) was used as a positive control.

Amrine C.S., Long J.L., Raja H.A., Kurina S.J., Burdette J.E., Pearce C.J, & Oberlies N.H. (2019). Engineering Fluorine into Verticillins (Epipolythiodioxopiperazine Alkaloids) via Precursor-Directed Biosynthesis. Journal of natural products, 82(11), 3104-3110.

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Cytotoxicity Assay for HT-29 Colon Cancer Cells

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Human colon cancer cells HT-29 were purchased from the American Type Culture Collection (Manassas, VA). The cell line was propagated at 37°C in 5% CO₂ in RPMI 1640 medium, supplemented with fetal bovine serum (10%), penicillin (100 units/ml), and streptomycin (100 μg/ml). Cells in log phase growth were harvested by trypsinization followed by two washings to remove all traces of the enzyme. A total of 5,000 cells were seeded per well of a 96-well clear, flat-bottom plate (Microtest 96®, Falcon) and incubated overnight (37°C in 5% CO₂). Samples dissolved in DMSO were then diluted and added to the appropriate wells (concentrations: 20 μg/mL and 2 μg/mL; total volume: 100 μL; DMSO: 0.5%). The cells were incubated in the presence of test substance for 72 hours at 37°C and evaluated for viability with a commercial absorbance assay (CellTiter 96® AQueous One Solution Cell Proliferation Assay, Promega Corp Promega) that measured viable cells. Survival percentage, based on microplate reader (Synergy Mx, BioTek) readings of absorbance at 490 nm, was expressed in percentage relative to the solvent (DMSO) control. One of the authors (Dr. Wei-Lun Chen) performed the bioassay and interpreted the results (Ren et al., 2017 (link)).

Henkin J.M., Sydara K., Xayvue M., Souliya O., Kinghorn A.D., Burdette J.E., Chen W.L., Elkington B.G, & Soejarto D.D. (2017). Revisiting the linkage between ethnomedical use and development of new medicines: A novel plant collection strategy towards the discovery of anticancer agents. Journal of medicinal plant research, 11(40), 621-634.

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In Vitro Cytotoxicity Assay for Cancer Cell Lines

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Human melanoma cancer cells (MDA-MB-435), human breast cancer cells (MDA-MB-231), and human ovarian cancer cells (OVCAR3) were purchased from the American Type Culture Collection (Manassas, VA). Each cell line was propagated at 37°C in 5% CO₂ in RPMI 1640 medium supplemented with FBS (10%), penicillin (100 U/ml), and streptomycin (100 μg/ml). Cells in log phase were harvested by trypsinization, followed by two washing cycles to remove all traces of enzyme. A total of 5,000 cells were seeded per well of a 96-well clear, flat-bottom plate (Microtest 96; Falcon) and incubated overnight (37°C in 5% CO₂). Samples dissolved in DMSO were then diluted and added to the respective wells. The cells were incubated in the presence of test substance for 72 h at 37°C and evaluated for viability with a commercial absorbance assay (CellTiter 96 AQ_ueous One Solution cell proliferation assay; Promega Corp., Madison, WI) that measured viable cells. IC₅₀ values are expressed in micromolar values relative to the solvent (DMSO) control.

Choules M.P., Wolf N.M., Lee H., Anderson J.R., Grzelak E.M., Wang Y., Ma R., Gao W., McAlpine J.B., Jin Y.Y., Cheng J., Lee H., Suh J.W., Duc N.M., Paik S., Choe J.H., Jo E.K., Chang C.L., Lee J.S., Jaki B.U., Pauli G.F., Franzblau S.G, & Cho S. (2019). Rufomycin Targets ClpC1 Proteolysis in Mycobacterium tuberculosis and M. abscessus. Antimicrobial Agents and Chemotherapy, 63(3), e02204-18.

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Cytotoxicity of Equisetum arvense Extract

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MDA-MB-435 (melanoma) and OVCAR3 (ovarian) cancer cells were purchased from American Type Culture Collection (Manassas, VA, USA). The cell lines were proliferated at 37 °C in CO₂ incubator in RPMI 1640 media, supplemented with 10% fetal bovine serum (FBS), streptomycin (100 µg/mL) and penicillin (100 U/mL). Cells in the log phase of growth were trypsinized followed by washing to remove all the traces of enzymes. Cells in the concentration of 5,000 were seeded in each well of 96-well flat-bottom plate (Microtest 96®, Falcon, USA) and the cells were incubated overnight at 37 °C in CO₂ incubator. The crude extract and fractions of E. arvense (250 and 500 µg/mL) dissolved in dimethyl sulfoxide were added to corresponding wells. Dimethyl sulfoxide at the concentration of 0.05% was taken as a control. The cells were incubated with test samples and control for 72 h at 37 °C and observed for viability with an absorbance assay kit (CellTiter-Blue®, Madison, WI, USA). The % cell viability was calculated using the following formula;.

% C e l l v i a b i l i t y = \frac{Absorbance of sample}{Absorbance of control} \times 100

Z.i.a.-.u.r.-.R.e.h.m.a.n., Gurgul A., Youn I., Maldonado A., Wahid F., Che C.T, & Khan T. (2022). UHPLC-MS/MS-GNPS based phytochemical investigation of Equisetum arvense L. And evaluation of cytotoxicity against human melanoma and ovarian cancer cells. Saudi Journal of Biological Sciences, 29(6), 103271.

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Cytotoxicity Screening of Cardiac Glycosides

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The cytotoxicity of the cardiac glycosides was screened against the human cancer HT-29, MDA-MB-231, MDA-MB-435, or OVCAR3 cell lines, with a procedure reported previously,^{27 (link),28 (link)} with the vehicle and paclitaxel used as the negative and positive control, respectively. Briefly, after log-phase-growth, cells were seeded in 96-well clear flat-bottomed plates (Microtest 96, Falcon) and treated with a test sample or paclitaxel (both dissolved in DMSO and diluted to different concentrations required) or the vehicle (DMSO) for 72 h. Viability of cells was evaluated using a commercial absorbance assay (CellTiter 96 AQueous One Solution Cell Proliferation Assay, Promega Corp, Madison, WI, USA), with the IC₅₀ values calculated from the vehicle control. For cytotoxicity testing against the H1299 human non-small cell lung cancer cell line, cell proliferation was assessed using the MTT proliferation assay kit (Cayman Chemical, Ann Arbor, MI, USA). Cells were seeded in each well of a 96-well plate and treated with the samples for 24 h followed by a treatment with MTT for 4 h. The medium was removed, and 100 μL Crystal Dissolving Solution were added to each well. The absorbance of the solution was measured at 570 nm, with IC₅₀ values calculated from the vehicle control.

Ren Y., Ribas H.T., Heath K., Wu S., Ren J., Shriwas P., Chen X., Johnson M.E., Cheng X., Burdette J.E, & Kinghorn A.D. (2020). Na+/K+-ATPase-Targeted Cytotoxicity of (+)-Digoxin and Several Semi-synthetic Derivatives. Journal of natural products, 83(3), 638-648.

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Cytotoxicity Screening of Compounds

Cited 1 time

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Compounds (1–18) were tested for cytotoxicity against the MDA-MB-435^{27 (link)} human melanoma (HTB-129, ATCC) and the HT-29 human colon cancer (HTB-38,ATCC) cell lines as described previously.^{26 (link),28 (link)} Briefly, the cell lines were propagated at 37 °C in 5% CO₂ in RPMI 1640 medium, which was supplemented with 10% fetal bovine serum, 100 units/mL penicillin, and 100 μg/mL streptomycin. The cells were harvested by trypsinization while in their log phase and washed twice to remove all traces of enzyme. In every well of a 96-well plate (Microtest 96, Falcon), about 5,000 cells were seeded and incubated overnight in 5% CO₂ at 37 °C. Tested compounds dissolved in DMSO were incubated with the cells for 72 h at 37 °C and evaluated for viability with a commercial absorbance assay (CellTiter 96 AQ_ueous One Solution Cell Proliferation Assay, Promega Corp, Madison, WI). IC₅₀ values are expressed in μM relative to the solvent (DMSO) control. Vinblastine was used as a positive control.

Alali F., El-Elimat T., Albataineh H., Al-Balas Q., Al-Gharaibeh M., Falkinham JO I.I.I., Chen W.L., Swanson S.M, & Oberlies N.H. (2015). Cytotoxic Homoisoflavones from the Bulbs of Bellevalia eigii. Journal of natural products, 78(7), 1708-1715.

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Cytotoxicity Evaluation of Compounds Against Cancer Cell Lines

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Human melanoma cancer cells (MDA-MB-435), human breast cancer cells (MDA-MB-231), and human ovarian cancer cells (OVCAR3) were purchased from the American Type Culture Collection (Manassas, VA, USA). The cell lines were propagated at 37 °C in 5% CO₂ in RPMI 1640 medium, supplemented with fetal bovine serum (10%), penicillin (100 units/mL), and streptomycin (100 μg/mL). Cells in log phase growth were harvested by trypsinization followed by two washings with PBS to remove all traces of enzyme. A total of 5000 cells were seeded per well of a 96-well clear, flat-bottom plate (Microtest 96, Falcon) and incubated overnight (37 °C in 5% CO₂). Samples dissolved in DMSO were diluted and added to the appropriate wells in triplicate (concentrations: 25, 4, 0.8, 0.16, and 0.032 μM; total volume: 100 μL; DMSO < 0.5%). The cells were incubated in the presence of test substance for 72 h at 37 °C and evaluated for viability with a commercial absorbance assay (CellTiter 96 AQueous One Solution cell proliferation assay, Promega Corp., Madison, WI, USA) that measured viable cells. IC₅₀ values are expressed in μM relative to the solvent (DMSO) control. Paclitaxel was used as positive control.

Sun M., Guo B., Xu M., Zhao M., Onakpa M.M., Wu Z., Burdette J.E, & Che C.T. (2021). (9βH)- and 17-Nor-Pimaranes from Icacina oliviformis. Journal of natural products, 84(4), 949-955.

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Cell Viability and Proliferation Assay

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Cells were seeded in 96-well clear, flat-bottom plates (Microtest 96, Falcon, Corning, NY, USA) at a density of 500 cells per well. Each plate was incubated overnight at 37 °C under 5% CO₂. R1881 and bicalutamide were dissolved in DMSO and diluted to the appropriate concentrations to a total volume of 100 μL and 0.1% DMSO per well. The cells were then incubated for at 37 °C. Cell viability was assessed by Celltiter-Blue assay kit (Promega, Madison, WI, USA) as instructed by the manufacturer. After treatment, cells were incubated with CellTiter-Blue^® reagent for 1 h at 37 °C. Fluorescence was measured on a BioTek Synergy 2 microplate reader (BioTek, Winooski, VT, USA) at 560/590 nM. For sulforhodamine B (SRB) assays, MOE cells were plated at 5 × 10³ cells/mL in a 96-well plate then treated and incubated for 5 days followed by colorimetric assay as described previously [35 (link)]. Absorbance at 505 nM was measured on a BioTek Synergy 2 microplate reader.

Colina J.A., Zink K.E., Eliadis K., Salehi R., Gargus E.S., Wagner S.R., Moss K.J., Baligod S., Li K., Kirkpatrick B.J., Woodruff T.K., Tsang B.K., Sanchez L.M, & Burdette J.E. (2021). Fallopian Tube-Derived Tumor Cells Induce Testosterone Secretion from the Ovary, Increasing Epithelial Proliferation and Invasion. Cancers, 13(8), 1925.

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