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icc [2011/02/01 13:01]
mthwin removed
icc [2011/02/01 11:48] (current)
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 +ICC- Immuno Fluorescent Staining in Cell Culture
  
- </​dict>​ +AntibodiesRabbit Anti ______ 
- </​array>​ +            Mouse Anti  ______
- </​dict>​ +
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 +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)
 +
 +  - Wash cell in 1x PBS -5min
 +  - Fix cells with 4% PFA@ RT -30min
 +  - Wash cells with 0.01% PBS-Triton X 100 -5min
 +  - Permeabilize cells with 0.1% PBS-Triton X 100 -10min
 +  - Wash cells with 0.01% PBS-Triton X 100 -5min
 +  - Block cells with 10% Normal Donkey Serum in 0.01% PBS-Triton X 100 -60min @ RT
 +  - Incubate with _______________mouse anti __________ and ______________rabbit anti __________ in 3% Normal Donkey Serum in 0.01% PBS-Triton X 100 -2-16hrs @ 4C
 +  - Wash cells with 0.01% PBS-Triton X 100 -5min
 +  - Incubate with _______________anti mouse __________ and ______________anti rabbit __________ in 3% Normal Donkey Serum in 0.01% PBS-Triton X 100 -2hrs@ RT
 +  - Wash cell in 1x PBS -5min
 +  - Wash cell in 1x PBS -5min
 +  - Wash cell in 1x PBS -5min
 +
 +- mount onto slide-
/var/www/html/dokuwiki/data/pages/icc.txt · Last modified: 2011/02/01 11:48 (external edit)