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Fluoresence Microscopy

fluoresence antibody secondary primary

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2 replies to this topic

#1 Roxana Z

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Posted 18 May 2016 - 04:41 AM



I made some slides for fluorescence microscopy, but...


I have a problem. I made the wrong dilution for primary antibody so I have a higher concentration than I needed. I put the right concentration of secondary antibody. Do you think this will influence too much the binding of the secondary antibody? Can I rely on the results?


Thank you

#2 bob1


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Posted 20 May 2016 - 09:06 AM

You can't rely on the results, too high a primary concentration might result in off-target binding.

#3 Andrea Fortina

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Posted 31 May 2016 - 07:44 PM

It depends on how high you dilute your primary antibody. There could be a range for the application, if within the range, that is OK. If it is too high, you'd better do another one. 

Andrea Fortina

Genius only means hard-working for all one's life.

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