RE: plant protoplast sorting

From: Roger Burger (rburger@miltenyibiotec.com)
Date: Fri Mar 08 2002 - 13:34:42 EST


Hello Nigel

Although not published in the last 3 years, this article offers a unique
approach to protoplast isolation.

J Biochem Biophys Methods 1994 Jul;29(1):83-6


Selection and enrichment of differentially labeled plant protoplasts.

Barth S, Goerlich R, Schnabl H.

Institut fur Landwirtschaftliehe Botanik, Universitat Bonn, Germany.

A fast and efficient method for the selection of plasmamembrane-marked
protoplasts via magnetic-associated cell separation (MACS) was developed.
Isolated mesophyll protoplasts from sunflower (Helianthus annuus L.) were
covalently labeled at their plasma membrane with fluoresceine isothiocyanate
(FITC) with an efficiency of greater than 95%. They were subsequently
coupled to specific monoclonal antibodies against FITC, which were
conjugated to magnetic beads, and mixed with unlabeled hypocotyl
protoplasts. About 95% of the marked mesophyll protoplasts were separated by
the MACS system with a purity of greater than 80%. Protoplasts embedded in
agarose showed normal dividing activities.


Best Regards
Roger

Roger A. Burger, PhD
Technical Marketing Scientist
Miltenyi Biotec Inc.
Auburn, CA
(530 )887-5351
www.miltenyibiotec.com


> -----Original Message-----
> From: Nigel Blackhall [mailto:Nigel.Blackhall@nottingham.ac.uk]
> Sent: Tuesday, March 05, 2002 1:19 AM
> To: Cytometry Mailing List
> Subject: Re: plant protoplast sorting
>
>
>
> Dear all,
>
> I am currently updating a review article: Flow Cytometry of
> Plant Cells in Encyclopedia
> of Cell Technology, ed Spier RE.
>
> If anyone has any articles which they feel are relevant and
> occurred in the last three
> years I would greatly appreciate receiviing the details.
>
> Thanking everyone in advance,
>
> Nigel
>
> Nigel Blackhall
> Senior Experimental Officer
> Plant Science Division
> School of Biosciences
> University of Nottingham
> Sutton Bonnington Campus
> Lougborough
> Leicestershire
> tel  +44 (0) 115 9515151 ext 18501
> fax +44 (0) 115 9516334
>
> mobile 07810 123410
>



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