研究目的
Describing the immunodetection of endogenous FOXO by confocal microscopy as a chemical tool to quantify FOXO expression levels, its cellular location, and its active/inactive forms with relevant antibodies.
研究成果
The protocol described is an accurate method to detect FOXO proteins and other transcription factors, serving as a valuable tool for validating the activity of new molecules and understanding disease mechanisms.
研究不足
The protocol's effectiveness may vary depending on the cell line used and the quality of antibodies. Additional cell viability assays and compound solubility tests are recommended for accurate results.
1：Experimental Design and Method Selection:
The study uses confocal microscopy for immunodetection of endogenous FOXO proteins in U2OS osteosarcoma cell line.
2：Sample Selection and Data Sources:
U2OS human osteosarcoma cell line is used.
3：List of Experimental Equipment and Materials:
Includes T75 culture flasks, antibiotics, growth media, trypsinization solution, microplates, PBS, glycine buffer, permeabilization/blocking solution, antibodies, nuclear staining solution, and imaging hardware and software.
4：Experimental Procedures and Operational Workflow:
Involves sample preparation, antibody staining, and imaging steps.
5：Data Analysis Methods:
Fluorescent signal is analyzed for FOXO expression levels and cellular location.
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