Introduction 1 5 6 10 11 12 13 14 15 16 12 12 13 13 17 18 1 a d a p i n c 19 20 7 20 23 Fig. 1 DNA immunization vs. peptide immunization  Materials and methods Cloning of expression vectors and cell transfections www.rzpd.de 7 1 4 4 24 6 Ssp Sca Genetic immunization and purification of antibodies 20 22 6 1 4 7 Immunofluorescence analyses 6 5 FACS analyses 5 Immunoprecipitation analyses 7 6 Immunoblot analyses 7 5 Results and discussion Use of transiently transfected CHO cells to assess the specificity and titres of antisera against purinoceptors and ecto-enzymes 2 1 2 1 Fig. 2 a b c  Panels b c left panels a c right panels  Table 1 Titres, specificities, and applications of antibodies against members of the families of P2X purinoceptors, CD39-like ENTPDs, and CD203-like ENPPs Antibody Host Titre Specificity FACS/IFT Western blot P2X family 9316 rb 1:800 1 +++ (+) RG2 rt 1:800 1 +++ ++ CR29 rb 1:1600 4 +++ ++ CR30 rb 1:1600 1 +++ ++ 116 rb 1:800 4   RG22 rt 1:1600 4 +++ +++ K1G rb 1:1600 7 +++ − RH23 rt 1:1600 7 ++ − Hano43 rt mAb 7 ++ − 3428 rb 1:1600 7 ++ − Calt rb 1:2000 1 − +++ Abc rb 1:2000 4 − +++ Al rb 1:5000 mP2X7 − +++ ENTPD family 5441 rb 1:1600 hENTPD2 +++ − RG10 rt 1:1000 h/mENTPD2 +++ nd RG10-B28 rt mAb hENTPD2 +++ nd 5446 rb 1: 400 hENTPD5 +++ − CR44 rb 1:6,400 mENTPD1 +++ +++ RG9 rt 1:1600 mENTPD1 +++ nd RG9-A59 rt mAb mENTPD1 +++ − CR45 rb 1:1600 mENTPD2 +++ + CR63 rb 1:1600 mETNTPD2 +++ nd CR46 rb 1:400 mENTPD5 +++ − CR47 rb 1:200 mENTPD6 +++ − ENPP family K2G rb 1:200 hENPP2 +++ nd 7619 rb 1:800 hENPP2 +++ nd CR65 rb 1:800 mENPP4 +++ nd CR66 rb 1:400 mENPP5 +++ nd CR67 rb 1:800 mENPP6 +++ nd 7620 rb 1:3200 rENPP3 +++ nd Monitoring cell surface expression of purinoceptors and ecto-enzymes by flow cytometry 3 1 1 3 Fig. 3 a b shaded histograms a dotted lines b red lines a  Some of the antisera recognize the denatured cognate antigen in Western-blot analyses 4 7 4 4 Fig. 4 a b u m h  Affinity purification of purinoceptors and ecto-enzymes 5 Fig. 5 u t 4  Conclusion 1 4 7 19 20