实验概要

The following protocol provides a method of immunofluorescence of general PBMC by Peprotech.

实验步骤

The following protocol used human PBMC that were isolated from whole human blood(Ficoll method) and plated onto fibronectin-coated coverslips in 10% FCS/DMEMsupplemented with pen/strep and L-glutamine. Cells were allowed to adhere at 37˚Cfor 1 hour before being fixed in 4% paraformaldehyde for 20 minutes at roomtemperature.

1. Fixed cells were permeabilized with 0.1% Triton X-100/PBS for 2 minutes before being washed extensively with PBS.

2. Coverslips were blocked with 1% BSA/PBS.

3. Slides were stained with the primary antibody (diluted in 1%BSA/PBS) at room temperature for 1 hour.

4. Cells were stained with the secondary antibodies for cytoskeleton and nuclei detection.

5. Coverslips were mounted onto glass slides with medium and imaged.