How To Permeabilize THP-1

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
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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.

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