Colonic lamina propria cells from wild type or Nod2−/− mice were labelled with Vybrant fluorescent dye (Molecular Probes, Leiden, The Netherlands) and added to the upper well of transwell plates (Fisher Scientific, Loughborough, UK) at 1-8×105 per well and chemokines were added to the bottom well (CCL2 (10ng/ml) and CCL2 (1ng/ml), both R and D Systems, Abingdon, UK). As a control, cells were also plated in the absence of chemokine. After incubating for 1h at 37°C cells in the bottom well were stained with CD11c antibodies and the total number of dye-labelled cells counted using an Olympus BX51 upright microscope, captured using a Coolsnap ES camera (Photometrics) through MetaVue Software (Molecular Devices).
Vybrant fluorescent dye
Manufactured by Thermo Fisher Scientific
The Vybrant fluorescent dye is a versatile labeling reagent used for a variety of biological applications. It is designed to provide bright, stable fluorescence when applied to cells, proteins, or other biological samples. The core function of the Vybrant dye is to enable the visualization and detection of labeled targets through fluorescence microscopy or flow cytometry techniques.
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Lab products found in correlation
2 protocols using vybrant fluorescent dye
1
Chemokine-Induced Transwell Migration
2
Chemokine-Induced Transwell Migration
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Colonic lamina propria cells from wild type or Nod2−/− mice were labelled with Vybrant fluorescent dye (Molecular Probes, Leiden, The Netherlands) and added to the upper well of transwell plates (Fisher Scientific, Loughborough, UK) at 1-8×105 per well and chemokines were added to the bottom well (CCL2 (10ng/ml) and CCL2 (1ng/ml), both R and D Systems, Abingdon, UK). As a control, cells were also plated in the absence of chemokine. After incubating for 1h at 37°C cells in the bottom well were stained with CD11c antibodies and the total number of dye-labelled cells counted using an Olympus BX51 upright microscope, captured using a Coolsnap ES camera (Photometrics) through MetaVue Software (Molecular Devices).
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