2 replies to this topic

[medchemgirl]

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.

[bob1]

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).

[medchemgirl]

thank you!!

bob1, on Jun 3 2009, 07:16 PM, said:

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).