Immunofluorescence permeabilisation buffer (0.25% Triton X-100) recipe
Opens PFA-fixed cells so antibodies can reach intracellular targets: PBS + 0.25% Triton X-100, 10 min at room temperature.
🧮 Volume calculator
| Component | Amount | Final concentration (1X) |
|---|---|---|
| 10% Triton X-100 | 1.25 mL | 0.25% |
| 1X PBS | to final volume |
Steps
- Add the 10% Triton stock to PBS and mix.
- Store at 4 °C (weeks).
- Apply to fixed, washed cells for 10 min at room temperature, then wash with PBS.
Good to know
- To preserve membranes (e.g. membrane proteins), use 0.1% saponin — it is reversible, so keep it in all later solutions.
- Methanol-fixed cells are already permeabilised.
FAQ
How do I choose the Triton concentration?
Within 0.1–0.5%: higher for nuclear targets, lower for cytoplasmic or cytoskeletal ones.
This is a widely used standard composition. Amounts depend on the hydrate form and purity of your reagents — check the formula weight (FW) on the bottle and follow your institution's safety rules and the manufacturers' instructions.