Cargando…
LeukoCatch, a quick and efficient tool for the preparation of leukocyte extracts from blood
BACKGROUND: Whole-protein extracts from peripheral blood leukocytes are ideal for basic and clinical research. However, lack of a simple preparation technique has limited the use of such extracts. The aim of this study is to develop a simple and easy system that can selectively obtain leukocyte extr...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2011
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3170244/ https://www.ncbi.nlm.nih.gov/pubmed/21849019 http://dx.doi.org/10.1186/1472-6890-11-9 |
_version_ | 1782211594391912448 |
---|---|
author | Okuzaki, Daisuke Kimura, Shoichi Yabuta, Norikazu Ohmine, Toshinari Nojima, Hiroshi |
author_facet | Okuzaki, Daisuke Kimura, Shoichi Yabuta, Norikazu Ohmine, Toshinari Nojima, Hiroshi |
author_sort | Okuzaki, Daisuke |
collection | PubMed |
description | BACKGROUND: Whole-protein extracts from peripheral blood leukocytes are ideal for basic and clinical research. However, lack of a simple preparation technique has limited the use of such extracts. The aim of this study is to develop a simple and easy system that can selectively obtain leukocyte extracts without hemoglobin. METHODS: A filter that captures the leukocytes but not RBCs was set at the bottom of a 10-mL medical syringe by sandwiching it between plastic stoppers. The capturing efficiency of leukocytes with this tool, called LeukoCatch, was examined using human macrophage cells (MONO-MAC-6). The abilities of LeukoCatch system to capture the leukocyte proteins and to remove the hemoglobin from RBCs were tested by western blot analysis using human blood samples. RESULTS: This study presents the development of LeukoCatch, a novel tool that allows the preparation of leukocyte extracts from blood samples within 3 min without centrifugation. Tissue-cultured human macrophage cells were tested to determine the optimal filter numbers and pass-through frequencies of LeukoCatch, which was then applied to 2-mL blood samples. Samples were passed 2~5 times through a LeukoCatch equipped with 5 filters, washed twice with phosphate-buffered saline for red cell removal, and leukocyte proteins were extracted with 0.5 mL of elution buffer. Western blot analysis of the purified extract indicated that more than 90% of hemoglobin was removed by the LeukoCatch and that the protein recovery rate of leukocytes was at least 4 times better than that of the conventional centrifugation method. CONCLUSION: We conclude that LeukoCatch is useful not only for diagnosis at the bedside but also for basic research using blood samples or tissue culture cells. |
format | Online Article Text |
id | pubmed-3170244 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-31702442011-09-10 LeukoCatch, a quick and efficient tool for the preparation of leukocyte extracts from blood Okuzaki, Daisuke Kimura, Shoichi Yabuta, Norikazu Ohmine, Toshinari Nojima, Hiroshi BMC Clin Pathol Technical Advance BACKGROUND: Whole-protein extracts from peripheral blood leukocytes are ideal for basic and clinical research. However, lack of a simple preparation technique has limited the use of such extracts. The aim of this study is to develop a simple and easy system that can selectively obtain leukocyte extracts without hemoglobin. METHODS: A filter that captures the leukocytes but not RBCs was set at the bottom of a 10-mL medical syringe by sandwiching it between plastic stoppers. The capturing efficiency of leukocytes with this tool, called LeukoCatch, was examined using human macrophage cells (MONO-MAC-6). The abilities of LeukoCatch system to capture the leukocyte proteins and to remove the hemoglobin from RBCs were tested by western blot analysis using human blood samples. RESULTS: This study presents the development of LeukoCatch, a novel tool that allows the preparation of leukocyte extracts from blood samples within 3 min without centrifugation. Tissue-cultured human macrophage cells were tested to determine the optimal filter numbers and pass-through frequencies of LeukoCatch, which was then applied to 2-mL blood samples. Samples were passed 2~5 times through a LeukoCatch equipped with 5 filters, washed twice with phosphate-buffered saline for red cell removal, and leukocyte proteins were extracted with 0.5 mL of elution buffer. Western blot analysis of the purified extract indicated that more than 90% of hemoglobin was removed by the LeukoCatch and that the protein recovery rate of leukocytes was at least 4 times better than that of the conventional centrifugation method. CONCLUSION: We conclude that LeukoCatch is useful not only for diagnosis at the bedside but also for basic research using blood samples or tissue culture cells. BioMed Central 2011-08-17 /pmc/articles/PMC3170244/ /pubmed/21849019 http://dx.doi.org/10.1186/1472-6890-11-9 Text en Copyright ©2011 Okuzaki et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Technical Advance Okuzaki, Daisuke Kimura, Shoichi Yabuta, Norikazu Ohmine, Toshinari Nojima, Hiroshi LeukoCatch, a quick and efficient tool for the preparation of leukocyte extracts from blood |
title | LeukoCatch, a quick and efficient tool for the preparation of leukocyte extracts from blood |
title_full | LeukoCatch, a quick and efficient tool for the preparation of leukocyte extracts from blood |
title_fullStr | LeukoCatch, a quick and efficient tool for the preparation of leukocyte extracts from blood |
title_full_unstemmed | LeukoCatch, a quick and efficient tool for the preparation of leukocyte extracts from blood |
title_short | LeukoCatch, a quick and efficient tool for the preparation of leukocyte extracts from blood |
title_sort | leukocatch, a quick and efficient tool for the preparation of leukocyte extracts from blood |
topic | Technical Advance |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3170244/ https://www.ncbi.nlm.nih.gov/pubmed/21849019 http://dx.doi.org/10.1186/1472-6890-11-9 |
work_keys_str_mv | AT okuzakidaisuke leukocatchaquickandefficienttoolforthepreparationofleukocyteextractsfromblood AT kimurashoichi leukocatchaquickandefficienttoolforthepreparationofleukocyteextractsfromblood AT yabutanorikazu leukocatchaquickandefficienttoolforthepreparationofleukocyteextractsfromblood AT ohminetoshinari leukocatchaquickandefficienttoolforthepreparationofleukocyteextractsfromblood AT nojimahiroshi leukocatchaquickandefficienttoolforthepreparationofleukocyteextractsfromblood |