Re: Well-liked 405 secondary antibodies

Posted by George McNamara on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Well-liked-405-secondary-antibodies-tp7589467p7589477.html

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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
>> [phone]
>>
>>
>>