Abstract | BACKGROUND: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) has recently been utilized to accurately detect the amyloid proteins of renal amyloidosis. The present study investigated the optimal procedures for analyzing samples by LCMS/MS, and the advantage of using this technique to diagnosis renal amyloidosis. METHODS: To detect amyloid proteins, laser microdissected glomeruli from AL (n = 13) or AA (n = 10) renal amyloidosis patients were digested and analyzed by LCMS/MS. To determine the best procedures for analyzing samples by LCMS/MS, we examined the suitability of tissue samples, frozen or formalin-fixed paraffin-embedded (FFPE), the number of dissected glomeruli required for analysis (2, 10, or 50 glomeruli), and the amount of trypsin with or without dithiothreitol (DTT). We additionally compared the detection of amyloid proteins between immunostaining and LCMS/MS. RESULTS: CONCLUSION:
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Authors | Michiko Aoki, Dedong Kang, Akira Katayama, Naomi Kuwahara, Shinya Nagasaka, Yoko Endo, Mika Terasaki, Shinobu Kunugi, Yasuhiro Terasaki, Akira Shimizu |
Journal | Clinical and experimental nephrology
(Clin Exp Nephrol)
Vol. 22
Issue 4
Pg. 871-880
(Aug 2018)
ISSN: 1437-7799 [Electronic] Japan |
PMID | 29372474
(Publication Type: Journal Article)
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Topics |
- Amyloidosis
(diagnosis)
- Animals
- Chromatography, Liquid
- Humans
- Japan
- Kidney Diseases
(diagnosis)
- Kidney Glomerulus
(pathology)
- Mice
- Microdissection
- Rabbits
- Tandem Mass Spectrometry
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