Intradermal DNA immunization: antisera specific for the membrane lectin MR60/ERGIC-53.
The trafficking of intracellular membrane proteins in Golgi apparatus, endoplasmic reticulum or intermediate compartment has not yet been fully elucidated. The human MR60/ ERGIC-53 and the rat p58 proteins are one such protein; and to study them in cell-free and in situ systems, high quality monospecific antisera are required. Highly specific antisera have been obtained after immunization of mice with plasmids containing a gene encoding either the full length or a truncated protein. The best results were obtained after intradermal injections of a plasmid encoding a truncated protein comprising both the luminal carbohydrate recognition domain and the stem down to a cysteine residue close to the C-terminal end, but neither the transmembrane nor the cytosolic domains. Such antisera have a very high titer and are very efficient tools to visualize the MR60 protein in situ or to selectively precipitate the MR60 proteins from a whole cell lysate.
Carrière
V
V
Glycobiologie, Centre de Biophysique Moléculaire, CNRS, Orléans, France.
Landemarre
L
L
Altemayer
V
V
Motta
G
G
Monsigny
M
M
Roche
A C
AC
D000818:Animals; D000918:Antibody Specificity; D019556:COS Cells / Q000276:immunology; D002463:Cell Membrane Permeability; D005260:Female; D006801:Humans; D007106:Immune Sera; D007114:Immunization / Q000379:methods*; D007271:Injections, Intradermal; D037102:Lectins / Q000235:genetics* / Q000276:immunology*; D037241:Mannose-Binding Lectins; D008565:Membrane Proteins / Q000235:genetics* / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D009154:Mutation; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D011233:Precipitin Tests; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
D007106:Immune Sera; C084004:LMAN1 protein, human; D037102:Lectins; C501458:Lman1 protein, rat; D037241:Mannose-Binding Lectins; D008565:Membrane Proteins; D011994:Recombinant Proteins