JLat 9.2 was treated for 14h with 10ng/ml TNF in combination with either 2.5μM SAHA or 3μM Prostratin. Cells were washed once with 200μl PBS and fixed with 3.9% Formaldehyde (Miltenyi Biotec Inc., CA, USA) for 10 min at room temperature (RT). Afterward, cells were centrifugated at 500 x g for 5 min and 4°C and permeabilized for 30 min on ice using ice cold permeabilization buffer A (Miltenyi Biotec Inc., CA, USA). Two additional washing steps with PBS were performed after cell permeabilization, and cell nuclei were then stained with 0.1mg/ml DAPI (Sigma-Aldrich, St. Louis, USA) for 5 min in the dark at RT. After two washes with PBS, 1.5 × 105 cells were seeded into each experimental well of a half-area glass bottom 96-well plate (Corning) and imaged by taking 36 images per well using a 40x objective.
Half area glass bottom 96 well plate
Manufactured by Corning
The Half-area Glass Bottom 96-Well Plate is a laboratory equipment designed for cell culture and microscopy applications. It features a glass bottom in half of the wells, allowing for improved optical clarity and enhanced imaging performance compared to standard polystyrene well plates.
Automatically generated - may contain errors
2 protocols using half area glass bottom 96 well plate
1
Analyzing Cellular Response to TNF and Epigenetic Modulators
2
Analyzing Cellular Response to TNF and Epigenetic Modulators
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JLat 9.2 was treated for 14h with 10ng/ml TNF in combination with either 2.5μM SAHA or 3μM Prostratin. Cells were washed once with 200μl PBS and fixed with 3.9% Formaldehyde (Miltenyi Biotec Inc., CA, USA) for 10 min at room temperature (RT). Afterward, cells were centrifugated at 500 x g for 5 min and 4°C and permeabilized for 30 min on ice using ice cold permeabilization buffer A (Miltenyi Biotec Inc., CA, USA). Two additional washing steps with PBS were performed after cell permeabilization, and cell nuclei were then stained with 0.1mg/ml DAPI (Sigma-Aldrich, St. Louis, USA) for 5 min in the dark at RT. After two washes with PBS, 1.5 × 105 cells were seeded into each experimental well of a half-area glass bottom 96-well plate (Corning) and imaged by taking 36 images per well using a 40x objective.
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