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 Immunofluorescence Positive signal requires long exposure time y? [View Printable]
nkm

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HI,

my having trouble with my positive staining on paraffin tissue with immunofluorescence..

The exposure time required is very high since the signal is soo dim... it takes a long time to take pics...
I'm not sure what the problem could be?.
I would really appreciate any help?
.........................

Posted Jun 27, 2007, 19:42 PM
Tracy

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Can you increase the concentration of your primary antibody (i.e. lower dilution of your antibody) to make the signal stronger?
.........................

Posted Jun 27, 2007, 23:57 PM
nkm

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Hi Tracy,

I'm just worried if increase the concentration if that would cause too much non specific staining but I can try it..
thanks
.........................

Posted Jun 28, 2007, 12:47 PM
a_wandering_1

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I agree with your concern about increasing the antibody concentration. this usually just makes antibody stick in places it shouldn't.

What fluor are you using? Labeled primary or secondary? Homemade or commercial?
.........................

Posted Jun 28, 2007, 15:40 PM