Dear colleagues,

I am isolating intracellular bacteria from epithelial cells but keep encountering host cell contamination. My protocol:

  • Lysis: PBS wash → 0.1% Triton X-100 (10 min at 37°C, 20 min at RT).
  • Purification: Centrifuge 800g (10 min, 4°C) → Collect supernatant → Centrifuge 16,100g (10 min, 4°C) → Wash 2–3× with PBS.
  • Storage: Pellet stored at -20°C.
  • Any tips to reduce host contamination? Alternative detergents, washes, or centrifugation steps?

    Thanks in advance!

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