Black walled 96 well plates

Manufactured by BD

Sourced in United States

Black-walled 96-well plates are a type of laboratory equipment used for various cell-based assays and other applications. The plates have 96 individual wells with black walls, which help to minimize light interference and improve well-to-well consistency in optical measurements.

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Lab products found in correlation

Glomax multi plate reader, by Promega (2 mentions) Apo one homogenous caspase 3 7 assay, by Promega (2 mentions) Probenecid, by Merck Group (1 mentions) Fluostar optima plate reader, by BMG Labtech (1 mentions) Caspase glo 3 7 reagent, by Promega (1 mentions) Alamarblue reagent, by Thermo Fisher Scientific (1 mentions) Mtt solution, by Merck Group (1 mentions) Living image software, by PerkinElmer (1 mentions) Ivis lumina 2, by PerkinElmer (1 mentions) Staurosporine, by Merck Group (1 mentions)

Close spelling variants of this product

Gelatin-coated black-walled 96-well plates 96-well black plates 96-well plates

5 protocols using black walled 96 well plates

Salmonella-Based β-Lactamase Translocation Assay

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The β-lactamase translocation assay was performed as previously described (18 (link)). Briefly, HeLa cells, seeded in black-walled 96-well plates (BD Biosciences), were infected with Salmonella containing pWSK29-Spec (7 (link)) encoding TEM1-tagged effectors (Table S2) at a multiplicity of infection (MOI) of 100. Infected cells were centrifuged at 500 g for 5 min and incubated for 60 min at 37°C and 5% CO₂. The culture medium was replaced with 100 μl of 3 mM probenecid (Sigma-Aldrich), 20 mM Hepes in HBSS (Gibco), and 20 μl CCF2-AM LiveBLAzer-FRET B/G Loading Kit (Invitrogen) and incubated at room temperature, in the dark until 3 h postinfection. The cells were washed before the fluorescence was measured using a FLUOstar Optima plate reader (BMG Labtech) with an excitation wavelength of 410 nm and emission wavelengths of 450 and 520 nm. Response ratios were calculated by first subtracting the average background fluorescence for both 450 and 520 nm wavelengths from the fluorescence reading for each sample. The ratio of fluorescence at 450 nm to fluorescence at 520 nm for each sample was then divided by the uninfected ratio of fluorescence at these wavelengths.

McDowell M.A., Byrne A.M., Mylona E., Johnson R., Sagfors A., Crepin V.F., Lea S, & Frankel G. (2019). The S. Typhi effector StoD is an E3/E4 ubiquitin ligase which binds K48- and K63-linked diubiquitin. Life Science Alliance, 2(3), e201800272.

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Bcr-TMP Cytotoxicity Evaluation

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Cell viability was determined by MTT assays. After incubating the cells for 24 h with Bcr-TMP, MTT solution (Millipore Corp., Bedford, MA, USA) was added and incubated for 4 h. The insoluble formazan product was then dissolved in DMSO and the absorbance was measured at 570 nm with a microplate photometer (MPP 4008, Mikrotek, Overath, Germany). Proliferation assays of ACC cells were carried out as follows. Approximately 4000 cells were seeded per well in black-walled 96-well plates (BD, Franklin Lakes, NJ, USA) and then treated with desired concentrations of Bcr-TMP or DMSO as control; 72 h later the cells were assayed with Alamar blue reagent (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s instructions. Apoptosis assays were carried out using approximately 8000 ACC cells per well of a white-walled 96-well plate (BD) and subsequently treated for 24 h with DMSO or Bcr-TMP. Apoptosis was then assayed with the Caspase-Glo 3/7 reagent (Promega, Madison, WI, USA).

Yusenko M.V., Biyanee A., Frank D., Köhler L.H., Andersson M.K., Khandanpour C., Schobert R., Stenman G., Biersack B, & Klempnauer K.H. (2021). Bcr-TMP, a Novel Nanomolar-Active Compound That Exhibits Both MYB- and Microtubule-Inhibitory Activity. Cancers, 14(1), 43.

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Optical Imaging of Fluorescent Probes

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Optical imaging was performed on a PerkinElmer IVIS Lumina II. For open filter measurements, an exposure time of 30 seconds was used with large binning and f/stop=1. For selective emission filter measurements, the exposure time was 60 seconds. The Lumina has 4 emission filters with wavelengths 515-575, 575-650, 695-770 and 810-875 nm. The heater plate was set to room temperature to minimize evaporation. Black walled 96-well plates (BD Biosciences) were filled with 200 μL of probe at the indicated activities and concentrations. For pH imaging, the pH of the solution was adjusted using 0.1 M acetate (pH 4-5.5), MES (pH 5.5-6.5), phosphate (pH 6.5-8), Trizma (pH 8-9), borate (pH 9-10) or carbonate (pH 10-11) buffers. For switching experiments, the pH was adjusted using 1M HCl or NaOH. For phantom experiments, a 1-cm³ acrylic phantom (Data Spectrum Corporation) was filled with ¹⁸F-FDG and imaged under the same conditions. Image quantification was performed using Living Image software (PerkinElmer) by drawing regions of interest (ROIs) around each well and determining the radiance. A radioactive decay constant was applied to all data except for pH switching experiments to account for the time between plating and imaging.

Czupryna J., Kachur A.V., Blankemeyer E., Popov A.V., Karp J.S, & Delikatny E.J. (2015). Cerenkov specific contrast agents for detection of pH in vivo. Journal of nuclear medicine : official publication, Society of Nuclear Medicine, 56(3), 483-488.

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Caspase 3/7 Activation in HUVECs

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HUVECs were transfected 48 h before the assay. Cells were transferred to black-walled 96-well plates (Falcon) 4 h before the assay and incubated with EBM or 200nM Staurosporine (Sigma Aldrich) for 4 h. Caspase 3/7 activity was assayed according to the manufacturer´s protocol for ApoOne^® Homogenous Caspase 3/7 Assay (Promega). Fluorescence was measured with Glomax Multi plate reader (Promega).

Gimbel A.T., Koziarek S., Theodorou K., Schulz J.F., Stanicek L., Kremer V., Ali T., Günther S., Kumar S., Jo H., Hübner N., Maegdefessel L., Dimmeler S., van Heesch S, & Boon R.A. (2022). Aging-regulated TUG1 is dispensable for endothelial cell function. PLoS ONE, 17(9), e0265160.

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Caspase 3/7 Activity Assay in HUVECs

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HUVECs were transfected 48 h before the assay. Cells were transferred to black-walled 96-well plates (Falcon) 4 h before the assay and incubated with EBM or 200nM
Staurosporine (Sigma Aldrich) for 4 h. Caspase 3/7 activity was assayed according to the manufacturer´s protocol for ApoOne® Homogenous Caspase 3/7 Assay (Promega).
Fluorescence was measured with Glomax Multi plate reader (Promega).

Gimbel A.T., Koziarek S., Theodorou K., Schulz J.F., Stanicek L., Ali T., Günther S., Kumar S., Jo H., Hübner N., Mäegdefessel L., Dimmeler S., Heesch S.v., & Boon R.A. (2022). Aging-regulated TUG1 is dispensable for endothelial cell function.

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