Looking for f-tractin-TdTomato

[lechristophe]

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Hi,

I'm looking for the F-tractin-TdTomato used by several labs to label actin
in living cells (I already have red Lifeact constructs). All publications
point to Michael Schell from USUHS as the source of this plasmid, citing
his 2009 MBC paper:

Johnson HW, Schell MJ.
Neuronal IP3 3-kinase is an F-actin-bundling protein:
role in dendritic targeting and regulation of spine morphology.
Mol Biol Cell. 2009 Dec;20(24):5166-80. doi: 10.1091/mbc.E09-01-0083
PubMed PMID: 19846664

However, I've been unable to find a working email adress for Dr Schell, and
couldn't ask him for the plasmid. Does anybody have this plasmid, or know
where I could find it?

Thanks a lot,

--
Christophe Leterrier
Researcher
Axonal Domains Architecture Team
CRN2M CNRS UMR 7286
Aix Marseille University, France

[Richard Mort]

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We are having problems preserving our signal in fixed mouse embryos. The
flurophores we are using are Cherry and Venus. They look lovely in live
tissue but as soon as we fix (8-12 hours 4% PFA) we lose virtually all
the Venus and a lot of the Cherry signal. Does anybody have a good
fixation protocol/method they can reccomend to minimise background and
best preserve the signal?

Kind regards
Richard

--
Dr Richard Mort
MRC Human Genetics Unit
MRC IGMM
University of Edinburgh
Western General Hospital
Crewe Road
Edinburgh.
EH4 2XU, UK

Tel: [phone]
Fax: [phone]


The University of Edinburgh is a charitable body, registered in
Scotland, with registration number SC005336.

[Eric Shelden]

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We run into this in zebrafish too. I believe a few of the fluorescent proteins
are more tolerant to fixation, EGFP is supposed to be good, but then you
may need to address autofluorescence. Less fixation using smaller pieces of
tissue can also help. Unfortunately, we often find that immunostaining with
antibodies against the fluorescent protein often provides the only
satisfactory result.
Good luck,
Eric S