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Immunofluorescence stopped working!


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#1 tadala

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Posted 02 May 2011 - 04:18 AM

I developed procedure for CK19 and Mammaglobine staining and I had gut results, no background, no unspecific staining in izotyp contorls (almost). After a month I performed the same experiment and know I've got almost no staining in positive control and high staining in only secondary Ab control. (I didn't mix slides!). I used fresh blocking serum (10% FBS) and now my positive staining looked better but secondary ab control gives strong signal (as strong as positive control). Previously I made this experiments many times and it was never like this time!
What can I chack? What could it be?

I would be grateful for any help/advise!

#2 bob1

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Posted 02 May 2011 - 04:06 PM

Could your secondary be contaminated with bacteria?

#3 tadala

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Posted 03 May 2011 - 10:26 PM

Could your secondary be contaminated with bacteria?


I try to check this. But it looks as usually.
Thank you.




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