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unusual FITC background staining


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#1 biochem06

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Posted 24 January 2017 - 07:52 AM

Hi

 

I'm trying to do ANA staining on HEP2 cells. I'm Using FITC at 1:1000 dilution. As you can see from the image there is a weird speckled pattern over the entire slide. On the secondary only control slide (no primary antibody added) there is none of this speckled staining. Does anyone know what would cause this? I've never seen non-specific staining like this before. this is just a screen shot of the image.

 

Cells were fixed with 4% PFA, incubated with 10% BSA 1 hour and then primary followed by washing in PBS-T. Incubated with secondary at 1 in 1000 for 1 hour room temp then washed with PBS-T

 

Many thanks!

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Edited by biochem06, 24 January 2017 - 08:28 AM.


#2 bob1

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Posted 24 January 2017 - 08:21 AM

No image attached...



#3 biochem06

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Posted 24 January 2017 - 08:28 AM

sorry that should be it now






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