Labeling of platelet surface glycoproteins with biotin derivatives

Fabrizio Fabris, Immacolata Cordiano, Mario Mazzuccato, Paola Pradella, Maria Luigia Randi, Antonio Girolami

Research output: Contribution to journalArticlepeer-review

Abstract

We describe a non-radioactive method for the labeling of platelet surface proteins, consisting of platelet protein biotinylation by means of N-hydroxysuccinimido-biotin (NHS-B) and biotin-hydrazide (H-B) ; NHS-B labels proteins amino residues while H-B binds to periodate-modified sialoglycoproteins. Washed platelets were biotinylated and protein bands were detected after SDS-electrophoresis and western-blot using avidin-peroxidase and luminol as substrate to enhance the signal which was then detected by X-ray film . Biotin-labeled platelet proteins were also immunoprecipitated with monoclonal antibodies against glycoproteins Ib and the IIb-Illa complex. The use of periodate induced biotinylation is the method of choice for labeling platelet surface glycoproteins while NHS-B also labels internal proteins.The sensitivity of this new procedure is similar to that obtained with radiolabeling techniques; biotinylation does not interfere with the antigenic properties of Ib and IIb-IIIa glycoproteins.

Original languageEnglish
Pages (from-to)409-419
Number of pages11
JournalThrombosis Research
Volume66
Issue number4
DOIs
Publication statusPublished - May 15 1992

Keywords

  • biotin labeled platelets
  • platelet surface glycoproteins

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Hematology

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