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Easier elution of urinary proteins preserved on nitrocellulose membrane with heating
Author(s) -
QIN WEIWEI
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.825.1
Subject(s) - elution , nitrocellulose , chemistry , chromatography , membrane , collodion , urine , biomarker , biomarker discovery , urinary system , membrane protein , biochemistry , proteomics , biology , gene , endocrinology
Biomarker is measurable changes associated with the disease. Urine collects changes of the body and it is a better source for biomarker research than blood which is kept almost constant by homeostatic mechanisms of the body. Urimem, adsorbing urinary proteins on a piece of membrane which can be dried and stored in a vacuum bag, was proposed in the lab. In general, this approach is simple, economical, environmentally friendly and space‐saving, which enables comprehensive and systematic collection of urine from all people. It can potentially be an important part of the medical record. The preservation of urinary proteins on a membrane plays a vital role in searching biomarker in urine, and the efficient elution of proteins preserved on the membrane help to lower the barrier for more applications. The original elution procedure relied mainly on vortexing for complete dissolve of nitrocellulose in organic solvent. In current procedure, we raised the temperature for easier nitrocellulose dissolve, and then we precipitated proteins with gentle rotating to keep nitrocellulose dissolved during the procedure. SDS‐PAGE and LC‐MS/MS analysis showed there was no degradation of proteins prepared by heat‐and‐elute procedure. Its overlapping rate of the proteins prepared in separate experiments was 92.6%. By semi‐quantitative spectral counting the quantities of 85.2% of identified proteins’ CV values were within 20%. And the heating elution procedure was much simpler and more efficient than the previous one. It helps more researchers to adopt the membrane preservation of urinary proteins. Support or Funding Information National Basic Research Program of China (973 Program)(Nos. 2012CB517605, 2013CB530805); 111 Project (No. B08007).