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Hi Karl,

Rabut et al 2004 Nat Cell Biol
(http://www.embl-heidelberg.de/ExternalInfo/ellenberg/homepage/pdfs/Rabut200
4,NCB6,1114.pdf) obtained 120 GFP rotavirus like particles from one of the
authors (Cohen) of Dundr et al 2002 (pubmed abstract
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Ab
stract&list_uids=12490157
<http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A
bstract&list_uids=12490157> ), who is also an author on the original paper
that there are 120 GFPs in the VLP: A. Charpilienne et al 2001 (JBC - free
text at http://www.jbc.org/cgi/content/full/276/31/29361)
<http://www.jbc.org/cgi/content/full/276/31/29361)> .

The Dundr et al paper also references a couple of papers using His-tagged
GFP bound to Ni-chelate beads and similar approaches.

In the absence of photobleaching (i.e. no oxygen) why not standardized
against fluorescein MESF?


George

                -----Original Message-----
                From:   Karl Garsha [mailto:[log in to unmask]]
                Sent:   Tuesday, May 03, 2005 7:56 AM
                To:     [log in to unmask]
                Subject:        GFP standards or purified GFP?

                Search the CONFOCAL archive at
                http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

                While I'm thinking of it, does anyone know of a source of
calibration
                standards for work with fluorescent proteins, or
alternatively a source
                of purified GFP for creating such a set of standards?  With
enough
                motivation I'm sure one could grow up a bunch of cells,
solublize and
                affinity purify the GFP, dialize it to high concentration,
assay the
                concentration, and then make a serial dilution for dot-blot
standards,
                but this would be a huge amount of work.  It would be nice
to be able to
                buy purified GFP, CFP, YFP etc.  With all the attention on
                high-throughput instrumentation and quantitative assays, I
would think
                that someone would be selling calibration standards for
fluorescent
                proteins.  Thanks in advance for any suggestions.
                Best,
                Karl

                --
                Karl Garsha
                Light Microscopy Specialist
                Imaging Technology Group
                Beckman Institute for Advanced Science and Technology
                University of Illinois at Urbana-Champaign
                405 North Mathews Avenue
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