Literature DB >> 30731387

Optimization of von Willebrand factor multimer analysis in vertical mini-gel electrophoresis systems: A rapid procedure.

Camila Martos Thomazini1, Rosangela de Paula Silva Soares2, Tania Rubia Flores da Rocha2, Ana Teresa Azevedo Sachetto1, Marcelo Larami Santoro3.   

Abstract

Von Willebrand disease (VWD) is a common cause of bleeding worldwide. Analysis of von Willebrand factor (VWF) multimer distribution (VWF:MD) is essential to properly classify and treat different types of VWD, and it is performed using a SDS agarose gel electrophoresis followed by Western blotting, a handmade technique that demands days to be completed and requires skillful execution. Aiming both to facilitate gel production and to shorten the preparation time, we developed an uncomplicated technique to provide agility in the analysis of VWF:MD, so that it can be easily accomplished in the routine practice of hemostasis laboratories. On that account, we used a commercial vertical mini-gel electrophoresis system for SDS-PAGE and a semi-dry transfer system, which allowed us to analyze VWF:MD of various samples in a period shorter than 12 h. This technique differentiated VWF:MD in human and animal plasmas under normal, congenital and acquired (experimental envenomation by Bothrops jararaca snake) conditions. This optimized method is cheap, rapid, reproducible, easy to be performed, and uses electrophoresis and Western blotting systems available in most laboratories. All these advantages encourage hemostasis professionals to use it in their routine practices. In order to facilitate the setup and accomplishment of the whole procedure step by step, videos were appended to the article.
Copyright © 2019 Elsevier Ltd. All rights reserved.

Entities:  

Keywords:  ADAMTS13; Bleeding; Diagnosis; Electrophoresis; Proteolysis

Mesh:

Substances:

Year:  2019        PMID: 30731387     DOI: 10.1016/j.thromres.2019.01.018

Source DB:  PubMed          Journal:  Thromb Res        ISSN: 0049-3848            Impact factor:   3.944


  3 in total

1.  Von Willebrand Factor Multimers and the Relaxation Response: A One-Year Study.

Authors:  Carlo Dal Lin; Laura Acquasaliente; Sabino Iliceto; Vincenzo De Filippis; Giuseppe Vitiello; Francesco Tona
Journal:  Entropy (Basel)       Date:  2021-04-10       Impact factor: 2.524

2.  Optimization of western blotting for the detection of proteins of different molecular weight.

Authors:  Ghanshyam D Heda; Lisa Shrestha; Sagarina Thapa; Shreya Ghimire; Diptika Raut
Journal:  Biotechniques       Date:  2020-04-14       Impact factor: 1.993

3.  Involvement of von Willebrand factor and botrocetin in the thrombocytopenia induced by Bothrops jararaca snake venom.

Authors:  Camila Martos Thomazini; Ana Teresa Azevedo Sachetto; Cynthia Zaccanini de Albuquerque; Vânia Gomes de Moura Mattaraia; Ana Karina de Oliveira; Solange Maria de Toledo Serrano; Ivo Lebrun; Katia Cristina Barbaro; Marcelo Larami Santoro
Journal:  PLoS Negl Trop Dis       Date:  2021-09-03
  3 in total

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