Re: Well-liked 405 secondary antibodies

Posted by Debora Olivier (Keller) on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Well-liked-405-secondary-antibodies-tp7589467p7589483.html

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Hiya,

I've been using dylight 405 on our SP8 confocal (and airyscan) and it works
really well (though I am using it with my strongest primary for instance
Nucleopores).
used in combination with 488 (Alexa or Dylight) and other red secondaries
My mounting medium is based on sorbitol (
https://www.nature.com/articles/s41598-018-32191-x).
I haven't tested it in mountings like prolong et al, I could test it in
vectashield.
I can send a picture if needed.
Cheers

Debora
Debora Olivier, PhD
Research Engineer Microscopy,
Group « Telomeres and Genome Organization »
CBI / LBCMCP
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On Fri, May 3, 2019 at 6:07 AM George McNamara <[hidden email]>
wrote:

> *****
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> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi Craig, Joe et al,
>
> as previously noted in another reply, BV421 = Brilliant Violet 421,
> works fine on 405 nm laser. Available from BD Biosciences (acquired the
> manufacturer, Sirigen, for the Brilliants), and BioLegend.
>
> We have a use with a CD## direct labeled primary antibody added to mouse
> leukocytes, imaged live on our Leica SP8 confocal microscope (HyD1, 2nd
> gen hybrid detector), including both time series (100 min) and Z-series.
>
> I note BD Biosciences now has BUVs (excite well ~350 nm), a few BB's
> (excite 488 nm) and one BYG (excite ~561 nm).
>
> I have suggested to a number of users that they move on from DAPI and
> Hoechst to BioLegend's Zombie NIR, excite ok ~640 nm, emission peak ~750
> nm ... BV421-antibody is narrow emission spectrum, bright, and has lots
> of antibodies ... DAPI and Hoechst a broad spectrum, one trick fluors.
>
> enjoy,
>
> George
>
>
> On 5/2/2019 4:47 PM, Craig Brideau wrote:
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > What ever happened to Brilliant Violet? I remember a bit of marketing
> about
> > it a while ago, but never got around to trying it myself. Did anyone get
> a
> > chance to give it a go?
> >
> > Craig
> >
> > On Thu, May 2, 2019 at 2:27 PM Joe Lebowitz <[hidden email]>
> wrote:
> >
> >> *****
> >> To join, leave or search the confocal microscopy listserv, go to:
> >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> >> Post images on http://www.imgur.com and include the link in your
> posting.
> >> *****
> >>
> >> 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
> >>
> >> --------------------------------
> >>
> >> Joe Lebowitz
> >> Ph.D. Candidate
> >> NINDS Pre-doctoral Fellow
> >> Khoshbouei Lab
> >> UF Department of Neuroscience
> >> (561) 504-3810
> >>
> >>
> >>
>