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Author Topic: hepatocyte culture immunofluorecence  (Read 6346 times)

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Offline riversound

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hepatocyte culture immunofluorecence
« on: February 26, 2006, 02:17:39 PM »
hi, i am totally new to cultured cell immunofluorescence. in my recent experiment on immunofluorescent staining on some rat hepatocyte primary culture, the nonspecific staining caused by secondary antibodies was very high as i perform negative controls by ommitting primary. i used both donkey anti mouse and anti rabbit seconday antibodies. i'd like to hear advices and suggestions. thanks a lot!
riversound

hepatocyte culture immunofluorecence
« on: February 26, 2006, 02:17:39 PM »

Offline richard03

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hepatocyte culture immunofluorecence
« Reply #1 on: February 27, 2006, 10:45:28 PM »
Did you block with normal serum raised from same species as secondary antibody? In your case, you should use normal donkey serum blocking since you are using donkey anti-mouse, rabbit secondary.

Some more detailed procedure will help.

Offline riversound

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hepatocyte culture immunofluorecence
« Reply #2 on: March 03, 2006, 06:31:07 PM »
yes, i blocked with 10% NDS for 30 min prior primary antibodies.

hepatocyte culture immunofluorecence
« Reply #2 on: March 03, 2006, 06:31:07 PM »