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  2. → Viewing Profile: Posts: bkbk

bkbk

Member Since 29 May 2012
Offline Last Active Sep 18 2012 04:09 PM
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Posts I've Made

In Topic: Unusual Non-specific Binding in Immunofluorescence

01 June 2012 - 08:23 AM

I decided to compare the particular antibody with another antibody that targets another viral protein, and I found that the latter gave me clean results in mock infected cells.  So in conclusion, something is wrong with the former antibody.

hopefully the new antibody i ordered last night will work

In Topic: Unusual Non-specific Binding in Immunofluorescence

30 May 2012 - 04:40 PM

yes i have tried titrating both the primary and secondary antibodies. i begin to lose my target protein signal. i even tried secondary conjugate by itself and I got no fluorescence. this suggests that something is wrong with the primary step. i even tried blocking with 10% Normal donkey serum in PBS-T today instead of 1%, and I still got background in my mock infected cells.

i also found that there were very small white/peachy mush in my stock primary antibody...is this a problem?

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