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Immunofluorescence Positive signal requires long exposure time y? [View Printable]
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nkm
Group: Member Posts: 12 Joined: Mar 29, 2007
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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?
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| Posted Jun 27, 2007, 19:42 PM |
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Tracy
Group: Moderators Posts: 232 Joined: Feb 09, 2006
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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?
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| Posted Jun 27, 2007, 23:57 PM |
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nkm
Group: Member Posts: 12 Joined: Mar 29, 2007
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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
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| Posted Jun 28, 2007, 12:47 PM |
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a_wandering_1
Group: Member Posts: 15 Joined: Jun 05, 2007
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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?
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| Posted Jun 28, 2007, 15:40 PM |
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