trans-Golgi retention of a plasma membrane protein: mutations in the cytoplasmic domain of the asialoglycoprotein receptor subunit H1 result in trans-Golgi retention

Antisera used for detection of ASGP receptor subunits were raised in rabbits against the purified human receptor (62-5) or against synthetic peptides corresponding to the carboxy-terminal sequence of either H1 (anti-H1C) or H2 (anti-H2C). For immunofluorescence staining, IgG from anti-H1C and anti-H2C were isolated using protein A--Sepharose. Rabbit polyclonal antibodies anti-human milk galactosyltransferase (Nll; from E. Berger, University of Ziirich), anti-bovine cation-independent M6P receptor (from B. Hoflack, EMBL), anti-rat TGN-38/41 (from G. Banting, University of Bristol), and anti-human aminopeptidase N (from O. Nordn and H. SjOstrSm, University of Copenhagen) were used as serum preparations. Mouse monoclonal antibodies anti-canine LAMP-1 (AC17; from E. Rodriguez-Boulan, Cornell University), and anti-bovine ~-adaptin (100:3; Sigma Immunochemicals, Buchs, Switzerland) were used as ascites preparations. Mouse anti-human c-myc antibody 9El0 was used as culture supernatant. Secondary antibodies fluorescein-conjugated anti-mouse IgG and anti-rabbit IgG, and rhodamine-conjugated anti-rabbit IgG were from Cappel (Malvern, PA).