How to choose secondary antibody? - (Dec/25/2017 )

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.

-Mad Researcher-

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.

-bob1-