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How to choose secondary antibody?

secondary antibody alexa fluor immunocytochemistry

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#1 Mad Researcher

Mad Researcher

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Posted 25 December 2017 - 08:55 AM

Hi,
 
I wanted to know how should I decide which secondary antibody should I go for? 
For primary, I used an anti-rabbit. I know that I have to use a secondary anti-rabbit, so I used a goat anti-rabbit. But, how do I decide if I should choose Alexa Fluor 488 or Alexa Fluor 555 goat anti rabbit?
 
Thanks.

Cheers,

Mad Researcher

#2 bob1

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Posted 26 December 2017 - 07:52 AM

Go have a look at the specifications for the fluorescence filters on your microscope. 555 will need a specific filter, as will 488. In my experience most will have a filters suitable for 488, 594 and 680, as these are spectrally distinct, so you don't get cross-talk between the channels when staining with more than one colour.







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