Edu assay edu staining proliferation kit

Mice were sacrificed at indicated ages. Tissues were dissected and fixed in 10% Formaldehyde/PBS for histological analysis. Decalcification was performed in EDTA for 2 weeks on a necessary basis, and then tissues were processed in paraffin. Sections were cut, stained with Hematoxylin and Eosin (H & E), or subjected to other analyses according to the standard protocols.
Immunofluorescence was performed according to the protocol described in the data sheets of the anti-acetylated histone 3 lysine 27 (H3K27Ac) antibody (# 8173, Cell Signaling Technology) and anti-acetylate histone 3 lysine 9 (H3K9Ac) antibody (#9649, Cell Signaling Technology). The fluorescent labeled secondary antibody (Alexa 568 anti-rabbit goat antibody, A-11011) was purchased from ThermoFisher Scientific.
EdU (5-ethynyl-2′-deoxyuridine) labeling assay was performed according to the instruction of EdU Assay / EdU Staining Proliferation Kit (ab219801, Abcam). Proliferating cells were labeled by injecting 10mg/kg BW EdU to mice intraperitoneally 2 hours before sacrifice. Tissues were fixed, paraffin-processed, and subjected to EdU staining and DAPI nuclear staining.

EdU staining was performed to assess cell proliferation using the EdU Assay/EdU Staining Proliferation Kit (cat no. ab222421, Abcam). Each well of the 24-well plate 6 × 10⁴ cells were plated into each well of the 24-well plate and then transfected with indicated plasmids. After 48 hours, the cells were cultured with 50 μM EdU reagent for 2 hours and fixed with 4% formaldehyde for 0.5 hour, followed by incubation with glycine (2 mg/mL) for 0.25 hour and 0.5% Triton X-100 for 0.33 hour to permeabilize. Next, the cells were incubated with Hoechst 33342 for nuclear staining. The percentage of EdU positive cells was assessed under a fluorescence microscopy (Olympus IX73, Japan).