fluorescent dyes

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cornillon
cornillon's picture
fluorescent dyes

I have to do a cytotoxicity assay with calcein (ex:485, em:530) and EthD-1 (ex:495, em:645), and I'd like to have another marker for my target cells that I can easily find, but I don't know which one will be ok for the emission wavelenght, and currently use to mark commercial antibodies.

Vadim Zinchuk
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Have you tried quantum dots?

Have you tried quantum dots? These are new and I was reading encouraging reviews.

cornillon
cornillon's picture
Vadim Zinchuk wrote:Have you

Vadim Zinchuk wrote:

Have you tried quantum dots? These are new and I was reading encouraging reviews.

I'm sure it could be something interesting, but I also think it must be something very expensive and I don't know if you can find it easily...
So I prefer something more common, like PE-Cy7 (I think an emission at 760nm could be ok), but I don't know if you can find easily an antibody (I don't know for wich antigen now, but something common in mice's cells) with this dye... or if there is another dye with approximately the same emission, but easier to find in conjugates.

Protoplast
Protoplast's picture
Quote:Vadim Zinchuk

Quote:

Vadim Zinchuk
Have you tried quantum dots? These are new and I was reading encouraging reviews.

Vadium, i would be careful suggesting quantumn dots. My laboratory does a lot of work with them, and they are not only quite expensive, but not as bright as they are alleged to be, difficult to work with on occasion, and while they do come in a varied assortments of spectra with very sharp emission/excitation peaks, They are also questionable in relation to cytotoxicity, I have heard that when using them in vivo they can be hazardous.
A common antibody in mouse cells? Wouldnt there be some MHC protein you could utilise in some way?

Fraser Moss
Fraser Moss's picture
If you ever need to select a

If you ever need to select a dye, fluorophore or filter set that is compatible for your assay, can I suggest you use the Curvomatic web site.

https://www.omegafilters.com/front/curvomatic/spectra.php?product_id=&submit.x=41&submit.y=6

You can select your dye or filter to see all combinations of fluorphores and filters.

You could buy the appropriate wavelength Alexa dye (http://probes.invitrogen.com/servlets/publications?id=150) conjugated to Streptavidin and label your target with a biotin conjugated secondary Ab, although I do understand that a 2ndry preconjucated to fluorohore is preferable.