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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
biochemistry, genetics and molecular biology
Proteomic approach to human kidney glomerulus prepared by laser microdissection from frozen biopsy specimens: Exploration of proteome after removal of blood-derived proteins
Proteomics - Clinical Applications, Volume 6, No. 7-8, Year 2012
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Description
Purpose: Abundance of blood-derived proteins in glomeruli prepared by laser microdissection from human kidney biopsy specimens has hampered in-depth proteomic analysis of glomeruli. We attempted to establish experimental platform for in-depth proteomic analysis of glomeruli by removal of blood-derived proteins from frozen biopsy samples. Experimental design: Frozen sections of biopsy samples were exposed to repeated PBS washes prior to laser microdissection to remove blood-derived proteins, and glomerular dissectants were analyzed by MS. The depth of proteomic analysis was evaluated by dynamic range of identified proteins and detection of low-abundance proteins. Results: Two times PBS washes of frozen sections effectively eliminated blood-derived proteins in laser-microdissected glomeruli and gave an increased number of identified proteins. Analysis of glomeruli from single specimens by a linear ion trap-Orbitrap mass analyzer generated nonredundant, high-confidence datasets of more than 400 identified proteins with high reproducibility, which attained to a considerable depth of the glomerulus proteome as revealed by a wide dynamic range and identification of low-abundance proteins. Conclusions and clinical relevance: Implementation of washing of frozen section with PBS successfully removed blood-derived proteins and resulted in an in-depth proteomic analysis of laser-microdissected glomeruli, suggesting applicability to clinical study. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Authors & Co-Authors
Yoshida, Yutaka
Japan, Niigata
Niigata University, Graduate School of Medical and Dental Science
Nameta, Masaaki
Japan, Niigata
Niigata University, Graduate School of Medical and Dental Science
Kuwano, Masayoshi
Japan, Ibaraki
National Institute of Biomedical Innovation
Zhang, Ying
Japan, Niigata
Niigata University, Graduate School of Medical and Dental Science
Xu, Bo
Japan, Niigata
Niigata University, Graduate School of Medical and Dental Science
Magdeldin, Sameh
Japan, Niigata
Niigata University, Graduate School of Medical and Dental Science
Egypt, Ismailia
Faculty of Veterinary Medicine
Cui, Zenyui
Japan, Niigata
Niigata University, Graduate School of Medical and Dental Science
Fujinaka, Hidehiko
Japan, Niigata
Niigata University, Graduate School of Medical and Dental Science
Japan, Niigata
National Hospital Organization Niigata National Hospital
Yaoita, Eishin
Japan, Niigata
Niigata University, Graduate School of Medical and Dental Science
Tomonaga, Takeshi
Japan, Ibaraki
National Institute of Biomedical Innovation
Yamamoto, Tadashi
Japan, Niigata
Niigata University, Graduate School of Medical and Dental Science
Statistics
Citations: 11
Authors: 11
Affiliations: 4
Identifiers
Doi:
10.1002/prca.201200016
ISSN:
18628346
e-ISSN:
18628354
Study Approach
Qualitative