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ICC- Immuno Fluorescent Staining in Cell Culture

Antibodies: Rabbit Anti Mouse Anti

Primary Antibody Made in Rabbit/ Mouse

Secondary Antibody Made in

In Cell Culture - Procedure on Pure Neuronal Primary Cell Culture - adjust accordingly- (done on coverslips in 24 well plates)

  1. Wash cell in 1x PBS -5min

  2. Fix cells with 4% PFA@ RT -30min

  3. Wash cells with 0.01% PBS-Triton X 100 -5min

  4. Permeabilize cells with 0.1% PBS-Triton X 100 -10min

  5. Wash cells with 0.01% PBS-Triton X 100 -5min

  6. Block cells with 10% Normal Donkey Serum in 0.01% PBS-Triton X 100 -60min @ RT

  7. Incubate with _mouse anti and rabbit anti in 3% Normal Donkey Serum in 0.01% PBS-Triton X 100 -2-16hrs @ 4C

  8. Wash cells with 0.01% PBS-Triton X 100 -5min

  9. Incubate with _anti mouse and anti rabbit in 3% Normal Donkey Serum in 0.01% PBS-Triton X 100 -2hrs@ RT

  10. Wash cell in 1x PBS -5min

  11. Wash cell in 1x PBS -5min

  12. Wash cell in 1x PBS -5min

- mount onto slide-

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