I stained a free-floating ~300uM brian slice using 4 different 1º antibodies (rabbit, chick, mouse, rat).

For the mouse primary, I mistakenly used an Alexa-350 secondary, when I should have used 594.

The microscope we have in lab cannot detect the 350 fluorophore, so I'm wondering if I can re-stain the slice with the anti-mouse 594 secondary.

Thanks for your advice !

[cross-posted in reddit/labrats]

  • $\begingroup$ Are they the same clone? You might have blocked all the binding sites so you'll get a poor/no/noisy signal with the 594. $\endgroup$
    – CKM
    Apr 13 '19 at 23:00
  • $\begingroup$ Thanks for your input ! They are both Alexa conjugated anti-mouse secondaries, so I'm guessing they use the same clone to make them. (?) $\endgroup$
    – gaelgarcia
    Apr 14 '19 at 3:52

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