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Online Journal of Veterinary Research©
Volume 26 (7): 537-545, 2022.
Conjugated polyclonal antibody mouse IgG3 subclass in rabbit.
Somayeh Dadashi, Jafar Majidi, Leili Aghebati Maleki, Jalal Abdolalizadeh, Behzad Baradaran, Zahra Moradi,
Sadeq Eivazi, Majid Ahmadi, Naeemeh Majidi
Tabriz University of Medical Sciences, Tabriz, Iran.
Dadashi S, Majidi J, Maleki LA, Abdolalizadeh J, Baradaran B, Moradi Z, Eivazi S, Ahmadi M, Majidi N., Conjugated polyclonal antibody mouse IgG3 subclass in rabbit, Onl J Vet Res., Volume 26 (7): 537-545, 2022. Monoclonal and polyclonal antibodies are used for indirect flow cytometric, ELISA and cytotoxicity assays. We report a polyclonal antibody against mouse IgG3 in rabbit. IgG3 was purified by sepharose beads conjugated with protein for affinity chromatography and its purity confirmed by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). We then injected New Zealand white rabbit with the IgG3 intramuscularly and intradermally. We further purified antibody in rabbit serum with ammonium sulfate precipitation and ion-exchange chromatography and SDS-PAGE. This antibody was conjugated with horse radish peroxidase for titer of rabbit anti-mouse IgG3 and HRP of 64000 determined by ELISA. Purity reached ~95% and optimum dilution ~1:10000.
Key words: Anti-IgG3, Chromatography, Mouse IgG3, Polyclonal Antibody, Protein A.