Epitope analysis and antibody preparation of human polypeptide:N-acetylgalactosaminyltransferase-2 protein

Hybridoma (Larchmt). 2010 Dec;29(6):547-9. doi: 10.1089/hyb.2010.0057. Epub 2010 Dec 6.

Abstract

In order to study the subcellular localization, secondary structure, and surface properties of ppGalNAcT2, we have developed polyclonal antibodies against ricin of ppGalNAcT2. As indicated by ELISA and Western blot analysis, these antibodies exhibit good reactivity and high specificity. We also found that the prevailing epitopes of ppGalNAcT2 exist in the RICIN domain. Furthermore immunofluorescence assay has helped to localize ppGalNAcT2 to Golgi apparatus. The results suggest that ppGalNAcT2 might take part in modifying glycoproteins in the Golgi complex, thereby laying a valuable foundation for further study of ppGalNAcT2 in tumor cell lines.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibody Formation / immunology*
  • Blotting, Western
  • Cell Line
  • Computational Biology
  • DNA Primers / genetics
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes / immunology*
  • Fluorescent Antibody Technique
  • Golgi Apparatus / metabolism
  • Humans
  • N-Acetylgalactosaminyltransferases / immunology*
  • Polypeptide N-acetylgalactosaminyltransferase
  • Reverse Transcriptase Polymerase Chain Reaction
  • Ricin / immunology*

Substances

  • DNA Primers
  • Epitopes
  • Ricin
  • N-Acetylgalactosaminyltransferases