Hi Mona,
I have an equivalent clone (H2B-mCherry) in AX2 just using the act5 locus
for the knock in. At the moment I can just offer you a cell line that
still has the the resistance marker (Hygromycin).
I am doing CRE loxing with this clone at the moment. So in two weeks time
there should also be a recombined out version available if you like to
have it.
Best, Peggy
> We have an AX3 clone with H2B-Cherry knocked into the rps30 locus (on the
> chr 2 duplication). The selectable marker has been recombined out. The
> expression is about as even as we have seen for any knock-in. It is
> useful. If you are in AX2, we currently have no equivalent knock-in, so
> Annette's vector might be best.
>
>
> ________________________________
> From: DICTY <[log in to unmask]> on behalf of Hellen
> Ishikawa <[log in to unmask]>
> Sent: 20 July 2017 21:25
> To: [log in to unmask]
> Subject: [DICTY]
>
> Hi
> You can try to use Dicty expressing mRFP-Histone look Annette
> Muller-Taubenbeger.
>
> Best regards,
> Hellen
>
>
> On Thu, Jul 20, 2017 at 8:59 PM, mona saad
> <[log in to unmask]<mailto:[log in to unmask]>> wrote:
> I have tried Hoechst 33342 in dicty living cells, it enters dicty but it
> was located in different localizations as if Hoechst was encapsulated in
> vesicules... In fact, I'm not working on nuclear dynamics but another
> different field, I'm just want to localize a specific ligand in dicty
> (nucleus, mitochondria) so I need to color the nucleus in living cells..
> Thank you
>
> On Thu, Jul 20, 2017 at 6:43 PM, Koonce, Michael P (HEALTH)
> <[log in to unmask]<mailto:[log in to unmask]>> wrote:
> Dear Mona,
> We’ve tried, and in our experience Hoechst 33342 does not work with live
> cells. There are a couple of GFP- tagged nuclear proteins available (e.g.
> Wolfgang Nellen’s GFP-CenH3 construct in the Dicty stock center) to image
> nuclear dynamics, but these take more effort to set up and limit other
> labeling strategies.
> Cheers,
> Mike Koonce
>
>
> From: DICTY
> <[log in to unmask]<mailto:[log in to unmask]>>
> on behalf of mona saad
> <[log in to unmask]<mailto:[log in to unmask]>>
> Reply-To: DICTY
> <[log in to unmask]<mailto:[log in to unmask]>>
> Date: Thursday, July 20, 2017 at 8:15 AM
> To:
> "[log in to unmask]<mailto:[log in to unmask]>"
> <[log in to unmask]<mailto:[log in to unmask]>>
> Subject: [DICTY]
>
>
> ATTENTION: This email came from an external source. Do not open
> attachments or click on links from unknown senders or unexpected emails.
>
> Bonjour,
> I would ask if someone have tried Hoechst 33342 on dicty living cells and
> if he found it accumulating in nucleus?
> thank you
>
>
>
>
> --
> Dr. Hellen Ishikawa-Ankerhold
> Ludwig-Maximilians-Universität München
> Walter Brendel Centre of Experimental Medicine
> Head of Multiphoton Microscope Core Facility
> Medizinische Klinik 1- Klinikum Großhadern
> AG Prof. Dr. Steffen Massberg
> Marchioninistr.27
> 81377 München
>
> Tel:+49(0) 089-2180-76585 (lab)
> +49(0) 089-2180-76581 (office)
> e-mail to: [log in to unmask]<mailto:[log in to unmask]>
>
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