Black microtiter plate

Manufactured by Corning

Sourced in United States

Black microtiter plates are laboratory products designed for use in various scientific experiments and assays. They provide a standard platform for conducting multi-well tests, enabling the efficient handling and analysis of multiple samples simultaneously. The black color of the plates helps to minimize background interference, enhancing the accuracy and sensitivity of the measurements.

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Lab products found in correlation

Synergy ht spectrophotometer, by Agilent Technologies (1 mentions) Synergy ht, by Agilent Technologies (1 mentions) Nickel nitrilotriacetic acid ni nta sepharose, by Solarbio (1 mentions) Deoxynivalenol don, by Merck Group (1 mentions) Ovalbumin (ova), by Sangon (1 mentions) Model 2475, by Waters Corporation (1 mentions) Sunfire c18 5 μm column, by Waters Corporation (1 mentions) Bovine serum albumin (bsa), by Sangon (1 mentions) Neo microplate reader, by Agilent Technologies (1 mentions) Trypan blue, by Thermo Fisher Scientific (1 mentions) Infinite m1000 microplate reader, by Tecan (1 mentions)

5 protocols using black microtiter plate

Measuring Pla Protease Activity

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Pla protease activity measurement was performed as previously described (40 (link)). In brief, bacteria were grown as described above for AI-2 analysis and collected at the time of maximal AI-2 level. Cultures were centrifuged, washed twice, and resuspended in PBS to obtain a final OD₆₀₀ of 0.1 using a spectrophotometer (SmartSpec 300; Bio-Rad). For each sample, 50-μl suspensions were added to wells of a black microtiter plate (Costar Corning, Inc., Corning, NY) in triplicate. The hexapeptide substrate dabcyl-Arg-Arg-Ile-Asn-Arg-Glu-(edans)-NH₂, synthesized on Sieber amide resin (45 (link)), was added to the wells at a final concentration of 2.5 μg/50 μl. The kinetics of substrate cleavage by Pla was measured every 10 min for 3 h by a fluorometric assay (excitation/emission wavelengths, 360/460 nm) at 37°C on a BioTek Synergy HT spectrophotometer (BioTek Instruments, Inc., Winooski, VT).

Fitts E.C., Andersson J.A., Kirtley M.L., Sha J., Erova T.E., Chauhan S., Motin V.L, & Chopra A.K. (2016). New Insights into Autoinducer-2 Signaling as a Virulence Regulator in a Mouse Model of Pneumonic Plague. mSphere, 1(6), e00342-16.

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Assay of Yersinia pestis Protease Pla Activity

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All tested cultures (WT CO92, its mutants, and the complemented strains) were plated on HIB agar plates from −80°C glycerol stocks and incubated at 28°C for 36 h. Single colonies of each culture were then re-plated on fresh HIB agar plates and incubated at 28°C or 37°C for 20–22 h. Bacteria from each plate were suspended and diluted in PBS to obtain absorbance values (OD₆₀₀) of 0.1 (5 × 10⁷ cfu/ml) and 0.5 (2.5 × 10⁸ cfu/ml) in a spectrophotometer (SmartSpec^Tm 300, Bio-Rad). For each strain, 50 μl suspensions were added to wells of a black microtiter plate (Costar Corning Inc., Corning, NY) in triplicate. The hexapeptide substrate (5.0 μg/μl) in dimethyl sulfoxide (DMSO) [38 (link)], was diluted to a final concentration of 2.5 μg/50 μl, and added to each well containing bacterial cells. Pla activity was then measured in a fluorometric assay (Excitation/Emission 360/460nm) at 37°C on BioTek Synergy HT spectrophotometer (BioTek Instruments Inc., Winooski, VT). The substrate (DABCYL-Arg-Arg-Ile-Asn-Arg-Glu (EDANS)-NH₂) was synthesized on Sieber amide resin [38 (link)]. The kinetics of substrate cleavage by Pla was measured every 15 min for 2 h. The increase in relative fluorescence units (RFU) as a function of time for WT CO92, its mutants, and the complemented strains was recorded and the kinetics of substrate cleavage plotted using Prism (GraphPad, La Jolla, CA).

van Lier C.J., Tiner B.L., Chauhan S., Motin V.L., Fitts E.C., Huante M.B., Endsley J.J., Ponnusamy D., Sha J, & Chopra A.K. (2015). Further characterization of a highly attenuated Yersinia pestis CO92 mutant deleted for the genes encoding Braun lipoprotein and plasminogen activator protease in murine alveolar and primary human macrophages. Microbial pathogenesis, 80, 27-38.

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Quantitative Mycotoxin Analysis Using Ni-NTA Sepharose

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Nickel-nitrilotriacetic acid (Ni-NTA) Sepharose was procured from Solarbio (Beijing, China). OTA, OTC, and fumonisin B₁ (FB₁) were obtained from Pribolab (Singapore). Aflatoxin B₁ (AFB₁) and zearalenone (ZEN) were purchased from Fermentek (Jerusalem, Israel). OTB was from Bioaustralis (Smithfield, NSW, Australia). Deoxynivalenol (DON) was from Sigma-Aldrich (CA, USA). Bovine serum albumin (BSA) and ovalbumin (OVA) were obtained from Sangon Biotech Inc. (Shanghai, China). The black microtiter plates were from Corning Inc. (NY, USA). The mouse anti-OTA monoclonal antibody mAb YK232 was obtained from Yikang Biotech Inc. (Haikou, China). The anti-alpha fetoprotein (AFP) Nb was prepared in our lab. The engineered BL21(DE3) strain of E. coli containing the vector Nb28-pET25b for Nb28 expression was prepared previously. All other chemicals and organic solvents were of reagent grade or better.
Fluorescence spectra and UV–vis spectra were obtained using a spectral scanning multimode reader SP-Max 3500FL (Flash Spectrum Inc., Shanghai, China). The HPLC system consisted of a Model E2695 pump, a Model 2475 fluorescence detector, and a SunFire C18 5 μm column (Waters, MA, USA).

Tang Z., Liu X., Wang Y., Chen Q., Hammock B.D, & Xu Y. (2019). Nanobody-based fluorescence resonance energy transfer immunoassay for noncompetitive and simultaneous detection of ochratoxin A and ochratoxin B. Environmental pollution (Barking, Essex : 1987), 251, 238-245.

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Luminescence Spectroscopy of Lanthanide-CPA Complexes

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Excitation and emission spectra of the CPA/lanthanide complexes were determined using a Neo microplate reader (BioTek, Winooski, VT, USA). All spectra were collected at 25 °C in MeOH/H₂O (9 + 1) using normal (not time resolved) luminescence. Test volumes were 300 µL. Experiments were conducted in black microtiter plates (Corning, Inc., Kennebunk, ME, USA). General settings for the instrument included a gain setting of 150, scanning in 1 nm increments, with 50 measurements per increment. Lamp energy was set to “low”, and distance of the optics above the microplate was set to 4.5 mm. The read speed was “normal”. Emission scans were collected over the range of 500 to 650 nm at an excitation of 290 nm. Excitation scans were collected over the range of 250 nm to 360 nm with monitoring emission at either 545 nm (Tb³⁺) or 615 nm (Eu³⁺). To demonstrate the enhancement, excitation and emission spectra were collected with and without CPA. The concentrations that were used were determined empirically and were selected to use as little of the reagents as possible while still yielding spectra with significant signal (5000 counts per second or higher) to allow for direct comparisons of sensitivity. For Eu³⁺ this was with EuCl₃ at 2.5 µM, with and without CPA at 25 µM. For Tb³⁺ this was with TbCl₃ at 0.25 µM with and without CPA at 1.5 µM.

, & Maragos C.M. (2018). Complexation of the Mycotoxin Cyclopiazonic Acid with Lanthanides Yields Luminescent Products. Toxins, 10(7), 285.

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Cytotoxicity Assessment of Vaa-Dis

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The in vitro cytotoxic potency of Vaa-Dis was evaluated using the PrestoBlue TM viability assay (Invitrogen, USA) essentially as described previously. 9 (link) Cells were trypsinized and counted by Trypan blue (Thermo Scientific, USA) exclusion assay 10 using a hemocytometer (Fortuna, Germany). Cells were seeded in 96-well plates (TPP, Switzerland) at a density of 5000 cells/well in 100 μL RPMI-1640 medium and left to attach overnight. Cells were then treated with different concentrations of Vaa-Dis (0.005; 0.05; 0.5; 5; 50; 500 nM) for 0, 24, 48, and 72 h. After the treatment, 10 μL of PrestoBlue TM was added to each well, and the plates were incubated for 30 min at 37 °C. Cell supernatants were transferred to black microtiter plates (Corning, USA) and fluorescence was measured at an excitation of 560 nm and emission of 590 nm on an Infinite M1000 microplate reader (Tecan, Switzerland) to determine metabolic activity of the cells. Wells containing only the cell culturing medium and PrestoBlue TM were used as blank reference standards. Experiments were performed in triplicate. The normalization of cell viability was calculated as the ratio of sample absorbance to control absorbance (cells in media without Vaa-Dis).

Latinović Z., Leonardi A., Petan T., Žlajpah M, & Križaj I. (2017). Disintegrins from the Venom of Vipera ammodytes ammodytes Efficiently Inhibit Migration of Breast Cancer Cells. Acta chimica Slovenica, 64(3).

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