Type-specific immunodetection of human heart fatty acid-binding protein with polyclonal anti-peptide antibodies.
In order to develop specific antibodies against human heart cytoplasmic fatty acid-binding protein (H-FABPc), four oligo-peptides of 15-20 amino-acids each and corresponding with different antigenic parts of the human H-FABPc molecule, were synthesized. Polyclonal antibodies against these synthetic peptides were raised in mice (Balb/C) and rabbits (Flemish giant). When tested in enzyme linked immunosorbent assays (ELISA, antibody-capture assay), antisera against three of the four peptides showed a high immunoreactivity with the synthetic peptide selected for immunization as well as with the native human H-FABPc. Some cross-reactivity with the other synthetic peptides was observed for the rabbit antisera but not for those from mice. Polyclonal antibodies against synthetic peptides can be applied for the specific detection of the native protein in biological preparations containing proteins that show a high degree of homology with the protein to be assayed.
Kleine
A H
AH
Department of Physiology, University of Limburg, Maastricht, The Netherlands.
Glatz
J F
JF
van Nieuwenhoven
F A
FA
Vallinga
M I
MI
Salden
M H
MH
Bosman
F T
FT
Boersma
W J
WJ
Zegers
N D
ND
van der Vusse
G J
GJ
D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D002352:Carrier Proteins / Q000032:analysis* / Q000276:immunology; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009206:Myocardium / Q000737:chemistry*; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D011817:Rabbits; D025521:Tumor Suppressor Proteins
D002352:Carrier Proteins; D000939:Epitopes; C107608:FABP7 protein, human; C082667:Fabp5 protein, mouse; C086936:Fabp7 protein, mouse; D000071438:Fatty Acid-Binding Protein 7; D050556:Fatty Acid-Binding Proteins; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D025521:Tumor Suppressor Proteins