Conjugation of R-Phycoerythrin to a Polyclonal Antibody and F (ab')2 Fragment of a Polyclonal Antibody by Two Different Methods

Author(s):

Jafar Mahmoudian , Mahmood Jeddi , Tehrani , Hodjattallah Rabbani , Ahmad Reza Mahmoudi , Mohammad Mehdi Akhondi , Amir Hassan Zarnani , Leila Balaei Goli , Mahdokht Babaei , Roya Ghods

Message:
Abstract:
R-Phycoerythrin (R-PE), a fluorescent protein from phycobiliprotein family, is isolated from red algae. Conjugation of antibodies to R-PE facilitates multiple fluorescent staining methods. In the present study polyclonal antibodies and polyclonal F(ab'')2 fragment antibodies were conjugated to R-PE by two different methods. The efficiency of the methods was evaluated using Immunocytochemistry (ICC) and Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE). In the first conjugation method, PE was attached to SMCC linker followed by conjugation of antibody to PE-SMCC. In the second method, SH groups were added onto R-PE molecule, while the antibody was attached to SPDP linker. Then, the antibody-SPDP molecule was conjugated to R-PE. Our results showed that the two conjugation methods did not have any abrogative effects on the antibody binding activity.
Language:
English
Published:
Avicenna Journal of Medical Biotechnology, Volume:2 Issue: 2, Apr-Jun 2010
Page:
87
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