Immunodetection of biotinylated lymphocyte-surface proteins by enhanced chemiluminescence: a nonradioactive method for cell-surface protein analysis. Anal Biochem 1992 Jul;204(1):220-6
Date
07/01/1992Pubmed ID
1514691DOI
10.1016/0003-2697(92)90165-4Scopus ID
2-s2.0-0026691177 (requires institutional sign-in at Scopus site) 100 CitationsAbstract
Biotinylation and radioiodination have been compared for labeling lymphocyte-surface proteins and the labeled proteins symmetrically immunoprecipitated with antibodies recognizing major lymphocyte markers such as the murine Thy-1, CD25 (the alpha subunit of the interleukin-2 receptor), CD45, and human CD2 glycoproteins. The detection of biotinylated proteins by enhanced chemiluminescence after transfer to nitrocellulose was found to be fast and as efficient as the detection of iodinated proteins by autoradiography. The vectoriality of cell-surface biotinylation was ascertained by two-dimensional electrophoresis of the cellular extract in which the major cytoplasmic proteins were not found biotinylated. This nonradioactive labeling procedure offers a convenient and efficient alternative to radiolabeling of cell surfaces for the biochemical analysis of extracellular domains of membrane proteins.
Author List
Meier T, Arni S, Malarkannan S, Poincelet M, Hoessli DAuthor
Subramaniam Malarkannan PhD Professor in the Medicine department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsAntigens, Differentiation
Biotin
Cell Membrane
Evaluation Studies as Topic
Humans
Iodine Radioisotopes
Luminescent Measurements
Lymphocytes
Lymphoma, T-Cell
Membrane Proteins
Mice
Mice, Inbred BALB C
Precipitin Tests
