Generation of rabbit antibodies against death ligands by cDNA immunization

Specificity problems, especially in immunoblot analysis, have been shown for several commercial antibodies raised against the death ligand Fas ligand (FasL) using conventional protein and/or peptide immunizations. In this work, we have optimized the development of rabbit antisera and isolated pAb ag...

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Bibliographic Details
Published in:Journal of immunological methods 2006-12, Vol.317 (1), p.12-20
Main Authors: Diestre, C., Martínez-Lorenzo, M.J., Bosque, A., Naval, J., Larrad, L., Anel, A.
Format: Article
Language:English
Subjects:
Animals
Antibodies - immunology
Antibody Specificity
AP03L/TWEAK
Apo2L/TRAIL
Biological and medical sciences
cDNA
DNA, Complementary - immunology
Fas Ligand Protein - genetics
Fas Ligand Protein - immunology
FasL
Flow Cytometry
Fundamental and applied biological sciences. Psychology
Fundamental immunology
HeLa Cells
Humans
Immunization
Immunoblotting
Jurkat Cells
Molecular immunology
Polyclonal antibodies
Rabbits
Techniques
TNF-Related Apoptosis-Inducing Ligand - genetics
TNF-Related Apoptosis-Inducing Ligand - immunology
Transfection
Tumor Necrosis Factors - genetics
Tumor Necrosis Factors - immunology
Vaccination - methods
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Summary:Specificity problems, especially in immunoblot analysis, have been shown for several commercial antibodies raised against the death ligand Fas ligand (FasL) using conventional protein and/or peptide immunizations. In this work, we have optimized the development of rabbit antisera and isolated pAb against the death ligands FasL, Apo2 ligand/TRAIL and Apo3 ligand/TWEAK by cDNA intramuscular immunization. This alternative approach has generated specific pAb in all three cases, which are useful for immunoblot purposes. The present data suggest that for the production of antibodies against certain glycosylated membrane proteins, cDNA immunization could be the method of choice.
ISSN:0022-1759
1872-7905
DOI:10.1016/j.jim.2006.08.018