http://confocal-microscopy-list.275.s1.nabble.com/Well-liked-405-secondary-antibodies-tp7589467p7589478.html
BD business model limitation ... can be dealt with. And the fluorescence
biotin-mAb ... ideally get 1:1 conjugation, but if a complex forms.
instead of full length IgG. Or VHH nanobodies.
> *****
> 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.
> *****
>
> Craig,
>
> You've probably heard some of this from me, and I know that George McNamara
> has also been impressed with and written of the Brilliant Horizon fluors
> from BD (BD owns this from Sirigen purchase).
>
> Here's a reference using Brilliant Violet 421 & Brilliant Violet 480:
>
> "Multilocus Imaging of the E. coli Chromosome by Fluorescent In Situ
> Hybridization."
>
https://www.researchgate.net/publication/319289637_Multilocus_Imaging_of_the_E_coli_Chromosome_by_Fluorescent_In_Situ_Hybridization>
> BV421 is approx. 20-40-fold brighter than AF405. BV480 is also approx.
> 25-fold brighter than CFP (depending on who is measuring...)
>
> The problem remains the availability of reagents, as BD has decided that
> their business model is better served by reserving these reagents for their
> custom-conjugated Ab's for flow.....But I believe that BD still entertains
> custom requests for their reagents.
>
> IMHO, directly conjugated, primary Ab's are the best way to leverage these
> fluors because they're so bright that they don't need amplification. This
> removes the problem of species specificity....Why wouldn't one try this???
>
> Here's a link to a presentation that illustrates the efficacy of this
> approach of filling the gap of violet & blue fluors. Given the headlong
> stampede towards increased multiplexing, I don't understand how these
> fluors could be overlooked:
>
https://www.chroma.com/5-channel-fluorescence-imaging-simplified>
> I hesitate using the "COMMERCIAL RESPONSE" qualifier, because Chroma
> Technology has no formal relationship with BD Biosciences and receives no
> credit for purchases of related products. Chroma Technology profits only
> from sales of filter sets which are used to image these fluors.
>
> Jeff
>
> *Jeff Carmichael*
>
> *Director of Marketing*
>
> *
[hidden email] <
[hidden email]> | 802-428-2528*
>
>
>
> On Thu, May 2, 2019 at 4:47 PM Craig Brideau <
[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.
>> *****
>>
>> 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
>>>
>>>
>>>