How To Permeabilize THP-1

THP-1 cells can be permeabilized with detergents.

THP-1 is a human acute monocytic leukemia cell line, which was originally obtained from a 1-year-old male infant. The cell line is used for scientific research, and can be permeabilized with a few simple steps. Permeabilization is an essential process if you want to detect intracellular antigens in your THP-1 cell line. Detergents, such as saponin, Triton X-100 and Tween-20, are common permeabilization reagents, which help make holes in the THP-1 cell membrane. The exact permeabilization time and concentration of the reagent depends on the experiment. Always follow the instructions in your specific protocol.

Things You'll Need

• Poly-L-lysine coverslips
• 10 percent formaldehyde
• Detergent, such as Triton X-100
• Phosphate-buffered saline
• Pipettes
• Pipette tips

Instructions

1. Culture the THP-1 cells on a poly-L-lysine coverslip at 10,000 cells per milliliter, or at a concentration recommended by your protocol.

2. Fix the cells with a 10 percent formaldehyde solution for one hour, or a time period recommended by your protocol. Optionally, fix the cells after permeabilization

3. Wash the cells with phosphate-buffered saline three times by dipping the coverslips into a PBS solution. .

4. Permeabilize the THP-1 cells with 0.5 percent Triton X-100 solution for 25 minutes, or 0.05 percent saponin solution for 20 minutes at room temperature. Make the 0.5 percent Triton X-100 solution by adding 500 microliters of 10 percent Triton X-100 to 9500 microliters of PBS. Make the 0.05 percent saponin solution by adding 500 microliters of 1 percent saponin solution into 9500 microliters of PBS.

5. Wash three times with PBS. The THP-1 cells are now permeabilized and ready for blocking and incubation with your antibody.

Tips & Warnings

• Always use good laboratory practices when working with human cell lines, formaldehyde and detergents.

References

• ATCC: THP-1 Product Description
• Infection and Immunity; Induction of Proinflammatory Responses in the Human Monocytic Cell Line THP-1 by Campylobacter jejuni; Michael A. Jones et al.
• Journal of Virology; Human Cytomegalovirus Chemokine Receptor Gene US28 Is Transcribed in Latently Infected THP-1 Monocytes; Patrick S. Beisser et al.