EGS (ethyleneglycol-bis-succinimidyl-succinate) Fixation

Date Published: July 18, 2023

EGS (ethyleneglycol-bis-succinimidyl-succinate) fixation (method to preserve microtubules and membrane bound antigens)

  1. Rinse cells with pre-warmed (37°C) PBS.
  2. Prepare EGS stock solution: dissolve 45 mg in 1 ml DMSO (75 mM). Note that EGS is highly unstable in water. Less than 1 minute should elapse between preparing EGS stock and transferring the cover slips into the solution.
  3. Dilute EGS stock in PBS to 10 mM.
  4. Drain off excess PBS from cells and immediately transfer them into EGS solution.
  5. Incubate (covered) at 37°C for 10 minutes.
  6. Rinse 3 times with PBS (include 100 mM glycine in 2nd wash).
  7. Permeabilize with 0.1 % Triton X-100 for 4 minutes at room temperature.

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