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Dot Blot question


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#1 medchemgirl

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Posted 03 June 2009 - 08:01 AM

I just developed a dot blot and I noticed that my spots for 1ry and 2ry antibodies are detected. At first I thought that these were like negative controls, but now that I saw my results I realized that the 1ry and 2ry spots should be detected since they were absorbed on the membrane. Am I correct? is this a way to know my antibodies work? Next time I will have to spot a negative control sample.

#2 bob1

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Posted 03 June 2009 - 04:16 PM

You are correct, they work because you then developed you blot - the primary was detected because you used the appropriate secondary, and the secondary worked because it is already conjugated to the detection system (e.g. HRP, AP).

#3 medchemgirl

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Posted 09 July 2009 - 11:24 AM

thank you!!

You are correct, they work because you then developed you blot - the primary was detected because you used the appropriate secondary, and the secondary worked because it is already conjugated to the detection system (e.g. HRP, AP).






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