An immunization method for generation of high affinity antisera against glucose transporters useful in immunohistochemistry.
Antipeptide antisera from unique amino acid sequences of proteins, predicted from their cDNA, are useful to study cellular distribution of these proteins, but such peptides often are poorly immunogenic. We describe a secondary immunization method with repeated intravenous administration of KLH-conjugated peptides to boost the immune response rapidly and transiently as high as 60-fold to peptides of low immunogenicity (glucose transporters, GLUT3 and GLUT4). Such antisera are suitable for immunocytochemistry with excellent anatomic detail. This method may be generally useful as it is reproducible and can yield progressive titer increases when repeated.
Van Bueren
A M
AM
Department of Cell Biology, Oregon Health Sciences University, Portland.
Moholt-Siebert
M
M
Begley
D E
DE
McCall
A L
AL
D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002531:Cerebellum / Q000166:cytology / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D051274:Glucose Transporter Type 3; D051275:Glucose Transporter Type 4; D007106:Immune Sera / Q000096:biosynthesis / Q000276:immunology*; D007114:Immunization / Q000379:methods; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins / Q000032:analysis / Q000276:immunology*; D009124:Muscle Proteins; D009132:Muscles / Q000378:metabolism; D009419:Nerve Tissue Proteins; D010455:Peptides / Q000138:chemical synthesis / Q000276:immunology; D011817:Rabbits / Q000276:immunology; D051381:Rats; D013569:Synapses / Q000378:metabolism
D051274:Glucose Transporter Type 3; D051275:Glucose Transporter Type 4; D007106:Immune Sera; D009004:Monosaccharide Transport Proteins; D009124:Muscle Proteins; D009419:Nerve Tissue Proteins; D010455:Peptides; C495689:Slc2a3 protein, mouse; C495690:Slc2a3 protein, rat; C495693:Slc2a4 protein, mouse; C495694:Slc2a4 protein, rat