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Anal Biochem. 2012 Jul 1;426(1):47-53. PubMed; doi 10.1016/j.ab.2012.03.026

Isolation of antibodies against different protein conformations using immunoaffinity chromatography.

Kuravsky, M. L.; Schmalhausen, E. V.; Pozdnyakova, N. V.; Muronetz, V. I.

A polyclonal antiserum obtained after the immunization of a rabbit with recombinant human sperm-specific glyceraldehyde-3-phosphate dehydrogenase lacking in 68 N-terminal amino acid residues (dN-GAPDS) was purified using different immunosorbents with immobilized dN-GAPDS in the native or denatured states. The procedure resulted in isolation of two types of polyclonal antibodies. The first type interacted with native recombinant dN-GAPDS as well as with native human sperm-specific glyceraldehyde-3-phosphate dehydrogenase, not cross-reacting with muscle glyceraldehyde-3-phosphate dehydrogenase (GAPD). The second type interacted with both native and denatured forms of the sperm-specific proteins, exhibiting some cross-reaction with GAPD. Thus, the suggested approach allows isolation of the antibodies against conformational or linear epitopes from the same polyclonal serum.



 


  Московский Государственный Университет имени М.В.Ломоносова



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Московского Государственного Университета имени М.В.Ломоносова


 





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