Immunoblotting Using Radiolabeled Reagents for Detection

Methods Mol Biol. 2015:1314:73-8. doi: 10.1007/978-1-4939-2718-0_9.

Abstract

Development of immunoblots is commonly performed using enzyme-labeled antibodies which convert soluble substrates into insoluble colored products. A simple, rapid, and sensitive alternative method which produces low background and allows a rapid quantitative evaluation is the use of radiolabeled antibodies or protein A conjugates. Here we describe the use of iodinated secondary antibodies for immunodetection of an autoantigen during HPLC purification.

MeSH terms

  • Autoantigens / analysis*
  • Autoantigens / isolation & purification
  • Blotting, Western / methods
  • Chromatography, High Pressure Liquid / methods
  • Electrophoresis, Polyacrylamide Gel / methods
  • Halogenation
  • HeLa Cells
  • Humans
  • Immunoblotting / methods*
  • Immunoconjugates / chemistry*
  • Indicators and Reagents / chemistry
  • Iodine Radioisotopes / chemistry*
  • Ribonucleoproteins / analysis*
  • Ribonucleoproteins / isolation & purification
  • SS-B Antigen
  • Staphylococcal Protein A / chemistry

Substances

  • Autoantigens
  • Immunoconjugates
  • Indicators and Reagents
  • Iodine Radioisotopes
  • Ribonucleoproteins
  • Staphylococcal Protein A