Re: Well-liked 405 secondary antibodies

Posted by Jacqueline Ross on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Well-liked-405-secondary-antibodies-tp7589467p7589472.html

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Hi Joe,

We also find that AlexaFluor 405 performs the best for our confocal systems as all except one have 405nm lasers. One other comment I would make is that depending on which filter set you have available for observation, the signal may look very weak to your eye (as it's violet) but may be much better than you think. I have often been pleasantly surprised by the signal strength of staining with this fluorophore when I switch over to confocal. I would also suggest increasing the concentration of the secondary and putting your best primary antibody into this channel if possible.

Cheers,

Jacqui
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From: Confocal Microscopy List [[hidden email]] on behalf of Joe Lebowitz [[hidden email]]
Sent: 03 May 2019 08:16
To: [hidden email]
Subject: Well-liked 405 secondary antibodies

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Hello all,

We have had some reproducibly disappointing results with Alexa Fluor 405
tagged secondary antibodies.

Does anyone have a go-to secondary in this color other than the AF? We are
trying to multiplex as many channels as possible and this is the only one we
struggle to get good brightness/SNR with.

Thanks!

joe

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Joe Lebowitz
Ph.D. Candidate
NINDS Pre-doctoral Fellow
Khoshbouei Lab
UF Department of Neuroscience
[phone]