Indeed, there are on average only 500 CR1 (CD35) molecules per human RBC, which doesn't really make a bright direct staining easy. I have been using home-conjugated mAbs for CD35 (therefore I don`t really know what is available on the market) and I got a good pos./neg. discrimination only with APC labelled mAbs (regardless of the specificity or avidity of the mAbs). Alternatively, you may try a biotinilated anti-CD35 + some kind of conjugated avidin. This would allow you to do multicolor analysis and should amplify the signal. Alessandra LUTI-IDM Institut Biomédical des Cordeliers 15, rue de l'Ecole de Médecine 75006 Paris tel. +33 (0)1 53 10 17 75 fax +33 (0)1 53 10 17 80 http://www.idm-biotech.com ---------- >De : "Jose A. Stoute" <stoutej@net2000ke.com> >À : Cytometry Mailing List <cytometry@flowcyt.cyto.purdue.edu> >Objet : Primary Conjugated antibodies against CD35 and CD55 >Date : Mer 26 avr 2000 5:29 > > > Dear group, I am trying to setup assays of direct fluorescent staining > of RBCs for CD35 and CD55. All the primary conjugated antibodies I have > tried for these antigens that are commercially available give a very dim > response, especially for CD35 which is known to have low expression on > RBCs. I have had good results using indirect fluorescence using a > secondary anti-mouse but this would preclude multicolor analysis, hence > we would like to switch to direct fluorescence. Any suggestions as to > which antibodies I could use or the approach? > > Thank you in advance for your assistance. > > Jose A. Stoute > > -- > Jose A. Stoute, MD > Unit 64109, Box 401 > USAMRU-Kenya > APO AE 09831-4109 > e-mail: stoutej@net2000ke.com > Nairobi Tel 254-2-729303, Fax 254-2-714592 > Kisumu Tel 254-35-22942, Fax 254-35-22903 > Electronic Fax Service: 1-630-214-2008, 1-917-463-0373, 1-917-477-6048 > >Received on Thu Apr 27 13:47:48 2000
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