anti-GFP antibody, rabbit serum Antibodies productsSearch results for: anti-GFP antibody, rabbit serum Antibodies
#32397084 
2020/05/08
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Positive Selection of Specific Antibodies Produced against Fusion Proteins.
A method for the positive selection of specific antibodies for target proteins expressed as fusion proteins for the production of antiserum is presented. As proof of concept, the fusion protein FLAG::His::GFP::His::FLAG was expressed in , purified, and used for the immunization of rabbits. The obtained serum was precleared via protein A affinity. A CusF::FLAG fusion protein was expressed in the periplasm of and purified. GFP without tags was also expressed in and purified via organic extraction. These proteins were then coupled to NHS-activated sepharose and used for the positive selection of Anti-GFP and Anti-FLAG antibodies. The obtained sera were tested for their specificity against different protein samples and fusion proteins in Western blots. A high specificity of the antibodies could be achieved by a single affinity chromatography step. In general, we advise to express the target protein with different tags and in different compartments for antibody production and affinity chromatography.Lukas Kramberger-Kaplan, Tina Austerlitz, Holger Bohlmann
