Urinary protein fraction in healthy subjects using cellulose acetate membrane electrophoresis followed by colloidal silver staining

Abstract

We previously reported a rapid and highly sensitive colloidal silver staining solution suitable for the cellulose acetate membrane. This method was useful for detecting even very small amounts of urinary protein. In the present study, we examined urinary protein fractions in healthy subjects, using cellulose acetate membrane electrophoresis (CAE) with a highly sensitive colloidal silver staining, in an attempt to determine the clinical relevance of urinary protein fractions. Sixty unconcentrated spot urine specimens were analyzed by CAE and calculated by densitometry. All of the samples were separated into five fractions by CAE. The mean +/- 1 SD of the percentage of five fractions was 28.37 +/- 8.51 in albumin, 4.30 +/- 4.19 in alpha1-globulin, 14.41 +/- 6.14 in alpha2-globulin, 19.45 +/- 7.10 in beta-globulin, and 33.46 +/- 8.24 in gamma-globulin. The albumin/globulin (A/G) ratio was 0.41 +/- 0.17. These six items and the concentrations of total protein, albumin, and beta-N-acetyl-D-glucosaminidase (NAG) did not significantly differ between males and females. NAG is the marker of tubulointerstitial nephropathy. The results suggest that there are no gender-dependent differences in the urinary protein fractions of healthy subjects.

The analysis and characterisation of immuno-unreactive urinary albumin in healthy volunteers

“…Other investigators have analysed urinary protein patterns by various electrophoretic methods including cellulose acetate membrane electrophoresis [21][22][23], SDS-PAGE [22], capillary electrophoresis, and two-dimensional electrophoresis followed by densitometry. These methods would not be expected to detect the immuno-unreactive albumin as this molecule appears unstable under denaturing and/or reducing conditions that are commonly utilised in these separation methods.…”

Rapid and sensitive electrophoresis of urinary protein clearly reveals the pathophysiological feature of renal diseases

“…Cellulose acetate membrane electrophoresis/colloidal silver staining method used in this study was sensitive enough to apply urine samples without concentration to electrophoresis, even if the protein level was as low as 20 mg/dL. It could also be used to see protein profiles of urine from healthy subjects, 12 and to detect M-protein in urine. 3 This method could be a great advantage for nephrologists to routinely obtain a profile of proteinuria at bedside.…”

Cellulose Acetate Membrane Electrophoresis Based Urinary Proteomics for the Identification of Characteristic Proteins

“…The renal tissue of patient A (b) shows tubular atrophy, granuloma formation, inflammatory cell infiltrate, and cast formation (1)(2)(3)(4). In addition, tubular atrophy, interstitial fibrosis, tubular dropout, and interstitial edema (5)(6)(7)(8) were observed in the tissue of patient B (c).…”