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cell confluency in IF

immunofluorescence cell confluency

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2 replies to this topic

#1 trangtrang



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Posted 04 September 2013 - 08:39 AM

Dear all, 


Actually, I performed the IF on control cells and cells transfected with siRNA. I don't know how much the protein was deleted. 

The problem is that signal in 'siRNA' cell is really strong, sounds specific. But in control cells, the signal is weaker weaker. The morphology seems the same. 


Because the number of cells on the coverslips of cell transfected is fewer (90%) in comparison with the number of control cells. 


If the quantity of antibody is the same, could the antibody react and fix more in a smaller number of cells? 


Sorry for this stupid question.


Thanks in advance.

#2 bob1


    Thelymitra pulchella

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Posted 04 September 2013 - 03:08 PM

It is highly likely that the amount of antibody is saturating despite the number of cells.   It is more likely that your cells are more confluent on the control and the antibody or fix is not penetrating as well, so the staining is weaker.

#3 trangtrang



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Posted 05 September 2013 - 04:34 AM

Thanks for your answer. Try another test and see how it is.

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