Sensitive Method for Detection and Semiquantification of Bence Jones Protein by Cellulose Acetate Membrane Electrophoresis Using Colloidal Silver Staining

Matsuda, Kazuyuki; Hiratsuka, Nobuo; Koyama, Takatoshi; Kurihara, Yoshitaka; Hasegawa, Osamu; Itoh, Yoshio; Shiba, Kiyoko

doi:10.1093/clinchem/47.4.763

Cited by 50 publications

(17 citation statements)

References 17 publications

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“…Urinary total protein concentrations of 14 samples were determined according to the calibration curve obtained by a silver dot blot assay, as described by Matsuda et al (1,2) and Hiratsuka et al (3). The values are shown in Table 1.…”

Section: Urinary Total Protein Concentrationmentioning

confidence: 99%

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Cellulose acetate membrane electrophoresis in the analysis of urinary proteins in patients with tubulointerstitial nephritis

Kubota¹,

Machii²,

Hiratsuka³

et al. 2003

Clinical Laboratory Analysis

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Urinary proteins from 14 patients with tubulointerstitial nephritis were analyzed by cellulose acetate membrane electrophoresis. Urinary total protein concentrations were measured, and urinary 15 proteins (prealbumin, albumin, alpha(1)-microglobulin, alpha(1)-antitrypsin, alpha(2)-macroglobulin, haptoglobin, retinol binding protein, transferrin, beta(2)-microglobulin, IgA, IgG, kappa- and lambda-light chains, cystatin C, and lysozyme) were identified by the use of a rapid and highly sensitive colloidal silver staining reagent suited for use with cellulose acetate membranes, as reported previously by Matsuda et al. (J Clin Lab Anal 15:171-174, 2001; Clin Chem47:763-766, 2001) and Hiratsuka et al. (J Clin Lab Anal 10:403-406, 1996). We also analyzed urinary total protein concentration and urinary protein fractions according to the presence of acute or nonacute interstitial nephritis. In addition, the relationship between urinary protein fraction and complications of interstitial nephritis was analyzed. The goal of this work was to find a useful index for the diagnosis of tubulointerstitial nephritis.

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Section: Urinary Total Protein Concentrationmentioning

confidence: 99%

“…In this study, a rapid and highly sensitive colloidal silver staining reagent suitable for use with cellulose acetate membranes, as reported by Matsuda et al (1,2) and Hiratsuka et al (3), was used. First, 15 urinary proteins in samples from patients with tubulointerstitial nephritis were identified, fractionated, and analyzed.…”

Section: Introductionmentioning

confidence: 99%

Cellulose acetate membrane electrophoresis in the analysis of urinary proteins in patients with tubulointerstitial nephritis

Kubota¹,

Machii²,

Hiratsuka³

et al. 2003

Clinical Laboratory Analysis

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“…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.…”

Section: Discussionmentioning

confidence: 99%

“…The diagnostic approach to renal diseases with the electrophoretic pattern of urinary protein on cellulose acetate (CA) membrane was well known worldwide. [1][2][3][4] The findings differentiate whether proteinuria was caused by glomerular or tubular injury, and had an impact on understanding the pathophysiology of renal diseases at bedside. However, this informative analytical method has not been used routinely.…”

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confidence: 99%

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

Sakatsume

Kubota²,

Ogawa

et al. 2007

Nephrology

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“…The sample was electrophoresed at a constant current of 0.7 mA/cm per membrane for 25 min in veronal buffer (60 mmol/L, pH 8.6). Silver staining was carried out using our previously-described method (5).…”

Section: Electrophoresis and Immunofixationmentioning

confidence: 99%

Analysis of an expanded width of albumin fraction by cellulose acetate membrane electrophoresis in IgA nephropathy urine before treatment

Machii

Matsuda

Hiratsuka

et al. 2003

Clinical Laboratory Analysis

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Cellulose acetate membrane electrophoresis with colloidal silver stain re-vealed that the width of the albumin fraction in IgA nephropathy (IgAN) urine before treatment was significantly expanded. This phenomenon was not shown in IgAN urine after treatment or in non-IgAN urine. There was a reverse correlation between the width of the albumin fraction and the albumin con-centration in IgAN urine. By immuno-fixation, Tamm-Horsfall protein (THP) was located in the same position as the albumin band in IgAN urine before treatment; however, in the urine of a healthy subject it was located in the same position as alpha(1)-globulin. By ELISA, the THP-albumin complex concentration in IgAN urine before treatment was significantly higher than in the other two diseases. The width of the albumin fraction and the sodium ion concentra-tion of the urine were significantly correlated. The THP/albumin ratio in IgAN urine before treatment was significantly higher than in the other two groups. This suggests that the characteristic expanded width of albumin found by immunofixation indicates a THP-albumin complex, and that the sodium concentration of urine is involved in the formation of this complex.

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Sensitive Method for Detection and Semiquantification of Bence Jones Protein by Cellulose Acetate Membrane Electrophoresis Using Colloidal Silver Staining

Cited by 50 publications

References 17 publications

Cellulose acetate membrane electrophoresis in the analysis of urinary proteins in patients with tubulointerstitial nephritis

Cellulose acetate membrane electrophoresis in the analysis of urinary proteins in patients with tubulointerstitial nephritis

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

Analysis of an expanded width of albumin fraction by cellulose acetate membrane electrophoresis in IgA nephropathy urine before treatment

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