Western: wrong 2. Ab- R MY RESULTS USELESS?! - (Oct/10/2006 )

Hello everybody! I have a BIIIG problem. I used a wrong second antibody (rabbit instead of mouse) and -of yourse- faced problems while developing... still...I got some bands. Which even fit my predictions. And they have the right size. So: are these results reliable? Is a cross-reaction possible? Or should I just forget about it? And start all over again?

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yes cross reaction is possible, however, why don't you incubate your membrane with the right antibody now? you would get nice bands.

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Do I have to incubate with the first Antibody again prior to incubating with the right secondary Ab? or follow a certain washing procedure?
Thank you!

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just wash the substrate, and incubate with the second antibody. the primary antibody is still bound, don't worry (unless all the blot strippers would have less problems !)

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Thank you Missele!!!

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you're welcome

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