Re: dynal beads- help

From: Andy Riddell (ar3@mrc-lmb.cam.ac.uk)
Date: Fri Jan 16 1998 - 06:12:04 EST


>Date: Thu, 15 Jan 1998 12:16:00 -0800 (PST)
>From: Mara Rocchi <M.ROCCHI@CGNET.COM>
>Subject: dynal beads- help
>To: Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu>
>
>
>
>Dear Flow Folks:
>
>does anybody have practical suggestions about assessment of purity of
>recovered cells after magnetic (positive) sort using Dynal beads?
>
>Beads are coated with rat anti mouse isotype-specific Ab.
>I have problem in detaching beads from cells (incomplete detachment ), in
>beads  intrinsic fluorescence (when beads are run without staining as
>control at the FACS) and in cross-reactivity between the GAM that I am
>suppose to use to quantify the purity (according to the manufacturer
>protocol) and the RAM.
>
>I will really appreciate all your ideas.
>
>Thanks in advance for the help
>Mara
>
>
> -------------------------------------------
>Dr. Mara Rocchi, DVM Ph.D.,
>Lab 4,
>International Livestock Research Institute
>Nairobi, Kenya
>Tel: 254-2-630743 ext. 4403
>Fax: 254-2-631499
>e-mail: M.Rocchi@cgnet.com


Hi Mara,

	Miltenyi Biotec do a magnetic bead separation kit called MACS. They have an
anti FITC antibody with a conjugated magnetic bead. All you would need to do is
stain up your cells with a rat anti mouse isotype-specific FITC antibody and
then come in with the anti FITC-Bead Ab. Run the separated cells on a FACS
machine to estimate percentage purity and the count the cells. I don't think
this bead will be compatible with your Dynal Bead system though, but maybe Dynal
have a similar Ab to Milenyi Biotec.

EUROPE:
Miltenyi Biotec
Friedrich-Ebert-Staße 68,
51429 Bergisch Gladbach,
Germany.
Tel: 02204-8306-0
Fax: 02204-85197

USA:
251 Auburn Ravine Rd.,
Suite 208,
Auburn,
CA 95603 - 9858.
Tel: (916) 888-8871
Fax: (916) 888-8925



Hope this helps....


Andy..


Andy Riddell
PNAC
MRC-LMB
Hills Road
Cambridge
UK

tel (0) 1223 402218
fax (0) 1223 412178
email ar3@mrc-lmb.cam.ac.uk
Andy Riddell
PNAC
MRC-LMB
Hills Road
Cambridge
UK

tel (0) 1223 402218
fax (0) 1223 412178
email ar3@mrc-lmb.cam.ac.uk



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