The EDU assay was performed using an EDU Kit (Beyotime), according to the manufacturer’s instructions. 5 × 104 cells/well Kyse150 and TE1 cells transfected with si-negative-control or si-NRP1 were seeded onto 12-well plates, irradiated with 4 Gy, and cultured for 48 h, and then, the medium was switched to fresh DMEM supplemented with EDU (50 mmol/L). The cells were then incubated for 2 h, followed by fixation, permeabilization, and EDU staining with Azide 488 (Beyotime). After staining cell nuclei with DAPI, five areas were selected randomly for photography and calculation. And the EDU-positive cells were identified using Zen 3.0 (blue edition) imaging software (Carl Zeiss).
Azide 488
Manufactured by Beyotime
Azide 488 is a fluorescent dye that can be used for labeling biomolecules. It has an excitation wavelength of 488 nm and an emission wavelength of 520 nm.
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Lab products found in correlation
2 protocols using azide 488
1
EDU Assay for Cell Proliferation
2
EdU Incorporation Assay for Proliferation
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Cells were plated on coverslips (Thermo Fisher Scientific) and incubated overnight. The cells were incubated with 10μM 5-ethynyl-2ʹ-deoxyuridine (EdU) for 2 h and stained with Azide 488 (Beyotime) after fixation with 4% formaldehyde. Coverslips were mounted with ProLong Diamond Anti-fade reagent with DAPI (Invitrogen). Gray level images were acquired under a laser scanning microscope (Axio Imager.Z2, Carl Zeiss Co. Ltd.).
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