>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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