sodium-dodecyl-sulfate and Proteinuria

The application of isoelectric focusing (IEF), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), two-dimensional electrophoresis (2-DE) and capillary electrophoresis (CE) for high resolution electrophoretic analysis of human urinary proteins is reviewed. In each case, the information is tabulated chronologically with details of sample preparation, electrophoretic system, detection method and clinical application. The text includes an historical perspective of the use of each method for urinalysis and a detailed review of the application of the methods to the investigation of renal disease, renal transplantation, Bence Jones proteinuria (BJP), diabetes mellitus, cadmium toxicity, nephrolithiasis and cancers of the urogenital tract.

Topics: Electrophoresis; Electrophoresis, Capillary; Electrophoresis, Gel, Two-Dimensional; Electrophoresis, Polyacrylamide Gel; Humans; Isoelectric Focusing; Proteinuria; Sodium Dodecyl Sulfate

We evaluated a new sodium dodecyl sulfate-agarose gel electrophoresis (SDS-AGE) for urinary protein analysis in patients with multiple myeloma (MM; n = 47; ages, 62 +/- 2 years, mean +/- SE). Abnormal proteinuria (mean = 1872 +/- 360 mg/24 h) was present in 95% of the samples; 75% of the patients had some sign of renal dysfunction (glomerular and/or tubular) according to their SDS-AGE pattern. A band suggesting Bence Jones proteinuria (BJP) was detected in 40 vs 33 specimens by routine AGE. Immunofixation identified BJP in 38 patients; the calculated sensitivity of SDS-AGE for BJP was 97%. Excellent correlation (P <0.0001) was obtained with routine AGE (r = 0.994) and immunonephelometry (r = 0.963) for light chain quantification. SDS-AGE allows easy evaluation of renal dysfunction and shows high sensitivity for BJP detection. In a specialized laboratory, it is useful for following the progress of MM patients through the semiquantification of BJP.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Bence Jones Protein; Electrophoresis, Agar Gel; Female; Humans; Immunoelectrophoresis; Immunoglobulin Light Chains; Male; Middle Aged; Multiple Myeloma; Nephelometry and Turbidimetry; Proteinuria; Reproducibility of Results; Sensitivity and Specificity; Sodium Dodecyl Sulfate

Proteome treatments with peptide libraries in view of reducing high-abundance proteins and increasing the concentration of rare species involve the adsorption on solid-phase material. Subsequent elution of captured proteins may not be fully effective except when sequences of eluting agents are used. The standard way utilized up to the present has been a three- to four-step, sequential elution system consisting of various agents mixed together such as urea, thiourea, CHAPS, sodium chloride, citric or acetic acid and some polar solvents such as ACN and isopropanol. Elution sequences produce distinct fractions adding to the burden of having to analyze all of them. An alternative, highly effective, single elution to reduce the workload is here reported for the first time, namely elution in boiling 10% SDS added with 3% DTE. This single step elutes almost quantitatively the adsorbed proteins, thus ensuring, for all practical purposes, a full recovery. This high efficiency is believed to be due to the fact that the SDS micelles bury the polypeptide chains within their hydrophobic core, thus shielding them from the surroundings and impeding accidental adsorption to surfaces. Suggestions for selecting the best method to eliminate the excess of SDS for further protein analysis are also evaluated. The merits and limits of this novel system are assessed and discussed.

Topics: Adsorption; Adult; Chemical Precipitation; Cholic Acids; Citric Acid; Combinatorial Chemistry Techniques; Electrophoresis, Gel, Two-Dimensional; Female; Guanidine; Humans; Hydrogen-Ion Concentration; Male; Peptide Library; Proteinuria; Proteome; Sodium Dodecyl Sulfate; Thiourea; Urea

In order to investigate the mechanism of urinary tract stone formation, we analyzed protein components in urine and the stone. Urinary proteins of healthy subjects and urolithic patients as well as protein components urinary tract stone of the urolithic patients were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Electrophoretic patterns of urinary proteins of the patients differed from those of healthy subjects after separating protein patterns into those larger than 66kDa or smaller than 30kDa. Protein constituents of urinary tract stone were mainly separated into 18 bands ranging from 26.8 to 143 kDa. Major bands among these 18 bands differed among stones from different patients. On western blotting, the developed intensities of Tamm-Horsfall protein (THP) were fainter than those of healthy subjects. Whereas intensities of albumin (ALB) were stronger than those of healthy subjects. Moreover, blotting patterns of THP of the patients on non-reducing SDS-PAGE were obviously broad. Thus, we suggest that analysis of fractionated urinary proteins or protein components of urinary tract stone may provide a tool for monitoring the prognosis or relapse in the patients.

Topics: Adult; Aged; Albumins; Albuminuria; Biomarkers; Electrophoresis, Polyacrylamide Gel; Female; Humans; Male; Middle Aged; Molecular Weight; Mucoproteins; Proteins; Proteinuria; Sodium Dodecyl Sulfate; Urinary Calculi; Uromodulin

To evaluate results of SDS-agarose gel electrophoresis (AGE) of urinary proteins for use in defining glomerular and tubulointerstitial derangements, investigate patterns of high-molecular-weight (HMW) proteins for differentiating among glomerular disorders, and assess low-molecular-weight (LMW) proteins as markers of severity of tubulointerstitial disease in dogs.. 49 dogs with increased serum creatinine concentrations or abnormal renal protein loss.. Urinary proteins were examined by use of SDS-AGE and differentiated on the basis of molecular weight. The HMW proteins (> or = 69 kd) were considered indicative of glomerular origin, whereas LMW proteins (< 69 kd) were of tubular origin. Renal specimens were examined by use of light microscopy. Glomerular and tubulointerstitial lesions were differentiated by use of the classification for the World Health Organization and semiquantitative grading, respectively.. Sensitivity of SDS-AGE was 100% for detection of glomerular lesions and 92.6% for tubulointerstitial lesions; specificity was 40% and 62.5%, respectively. Although HMW urinary proteins were not significantly associated with the type of glomerular lesion, LMW urinary proteins were significantly associated with the grade of tubulointerstitial damage. Detection of 12- or 15-kd proteins or both was highly indicative of a severe tubulointerstitial lesion.. SDS-AGE of urinary proteins in dogs represents a noninvasive test with high sensitivity for identifying glomerular and tubulointerstitial damage, but low specificity limits its validity as a stand-alone test to differentiate between glomerular and tubulointerstitial lesions. The test is particularly useful for identifying dogs with advanced tubulointerstitial disease but cannot be used to characterize glomerular disorders.

Topics: Animals; Creatine; Dog Diseases; Dogs; Electrophoresis, Agar Gel; Kidney Diseases; Kidney Glomerulus; Proteinuria; Sensitivity and Specificity; Sodium Dodecyl Sulfate

Topics: Aminoglycosides; Anti-Bacterial Agents; Coloring Agents; Indicators and Reagents; Molybdenum; Proteins; Proteinuria; Pyrogallol; Sodium Dodecyl Sulfate

Influence of protein composition on total urinary protein assays was evaluated for pyrogallol red-molybdate both with and without sodium dodecyl sulfate (SDS) and pyrocatechol violet-molybdate complex (UPRO) techniques. Using mixtures of albumin and gamma-globulins (n = 8; albumin/globulin ratio, 0 to 10), mean recoveries were 79, 77, and 81% for pyrogallol red, pyrogallol red-SDS, and UPRO, respectively. Using diluted myeloma sera (n = 26; A/G ratio, 0.39 to 2.35), mean recovery by the UPRO method was 115% (vs. 63% for pyrogallol red and 83% pyrogallol red-SDS). Results positively correlated with A/G ratio for UPRO (r = 0.69; P < 0.001), pyrogallol red (r = 0.48; P < 0.05), but not pyrogallol red-SDS (r = 0.191; NS). The difference between UPRO and pyrogallol red assays correlated with the A/G ratio (r = 0.82; P < 0.001). In light chain proteinuria (n = 10), no significant difference (< 15%) was observed between techniques, whereas in glomerular selective proteinuria (n = 10), values were significantly higher with the UPRO assay (2.20 +/- 1.61 vs. 1.43 +/- 1.10 g/L; P < 0.02). Our results support the idea that screening for renal diseases can be performed with UPRO or pyrogallol red assays. However, since A/G ratios may vary with renal disease evolution, follow-up of patients with positive proteinuria should be performed using the same assay, preferably the pyrogallol red-SDS.

Topics: Bence Jones Protein; Benzenesulfonates; Coloring Agents; gamma-Globulins; Humans; Molybdenum; Multiple Myeloma; Proteinuria; Pyrogallol; Sensitivity and Specificity; Serum Albumin; Serum Albumin, Bovine; Sodium Dodecyl Sulfate; Solutions

Balkan nephropathy (BN) is an endemic disease, which leads to end-stage renal failure and artificial renal replacement therapy. Pathologically it is characterized by progressive interstitial nephritis in a large population of villages situated in the proximity of a bend of the Danube up to a distance of 100 km from the river in several parts of Bulgaria, Romania, and the former Yugoslavia. The urinary proteins of 19 patients with BN from the region of Vratza, Bulgaria were examined using ultrathin layer sodium dodecyl sulfate (SDS) pore-graduated polyacrylamide gel electrophoresis (PAGE) and silver staining. The documentation of urinary proteins pattern was performed using laser densitometry and consecutive electronic processing for the purpose of characterizing and quantifying protein excretion. Our results show that the proteinuria of BN is predominantly tubular, consisting of low molecular weight species (10-65 kilodaltons). The amount of tubular protein changes with the progression of the disease. SDS-polyacrylamide gel electrophoresis (PAGE) is a diagnostic method for early diagnosis of tubular failure in BN. Using our method of SDS-PAGE, tubular failure can be detected even at a total protein concentration below 0.1 g/L and when the serum creatinine concentration is normal. Additionally, our method of SDS-PAGE supports the differentiation of BN from glomerular disease.

Topics: Aged; Aged, 80 and over; Balkan Nephropathy; Blood Proteins; Coloring Agents; Creatinine; Densitometry; Diagnosis, Differential; Electrophoresis, Polyacrylamide Gel; Female; Humans; Kidney Failure, Chronic; Kidney Glomerulus; Kidney Tubules; Lasers; Male; Middle Aged; Molecular Weight; Proteins; Proteinuria; Regression Analysis; Renal Replacement Therapy; Silver; Sodium Dodecyl Sulfate; Surface-Active Agents

The patterns of urinary proteins in rats of different ages were examined on SDS gradient polyacrylamide gel electrophoresis coupled with silver staining. Proteins were fractionated into at least 26 bands. Densitometric measurements were used to characterize protein excretion patterns. The results showed that proteinuria in newborn, young and adult rats is predominantly tubular, consisting of low molecular-weight species. Conversely, late adults and old rats had a mixed glomerular pattern, with a steadily increasing excretion of albumin, IgG and transferrin, as was the case of other high molecular-weight proteins. Fragments of both immunoglobulins and albumin were found in all urine samples assayed. In 1 month old rats the percentage of Tamm-Hörsfall (T-H) protein was higher (P < 0.01) than in the remaining groups studied. In newborns, relatively high albumin, IgG and transferrin percentages were detected, as well as an alpha 1-acid glycoprotein and carbonic anhydrase excretion (P < 0.05 and P < 0.01 respectively) higher than that observed in the other age groups studied.

Topics: Aging; Albuminuria; Animals; Animals, Newborn; Carbonic Anhydrases; Electrophoresis, Polyacrylamide Gel; Immunoglobulin Fragments; Male; Mucoproteins; Orosomucoid; Proteinuria; Rats; Rats, Wistar; Silver; Sodium Dodecyl Sulfate; Staining and Labeling; Transferrin; Uromodulin

The aim of this study was the determination of the electrophoretic patterns and evaluation of the proteinuria type in the normal pregnancy and in women with gestational hypertension with or without pathological proteinuria, using high performance electrophoresis in SDS-polyacrylamide as analytical tool. The study was done with patients from the General Hospital of the Ministry of Health in Durango, México. Three groups were formed; one included normal pregnant women (n = 13); the second one was formed by patients with gestational hypertension without pathological proteinuria (n = 25); the third group was formed by patients with gestational hypertension and pathological proteinuria (n = 12). The electrophoretic pattern in the three groups showed high and low molecular weight proteins, corresponding to mixed proteinuria (glomerular and tubular). The samples from the group of hypertension of pregnancy with pathological proteinuria showed greater IgG and transferrin content. It is concluded that physiological proteinuria in the normal pregnancy and pathological proteinuria in gestational hypertension differ quantitatively in an absolute and relative manner. The analytical method employed in this study is useful in the evaluation of renal compromise in diseases coursing with proteinuria.

Topics: Adolescent; Adult; Diagnosis, Differential; Electrophoresis; Electrophoresis, Polyacrylamide Gel; Female; Humans; Pre-Eclampsia; Pregnancy; Proteinuria; Sodium Dodecyl Sulfate

In rats, urine has been identified as a major source of the allergens that cause laboratory animal allergy, an important occupational health problem.. Urinary proteins and allergens of Wistar rats were studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. Proteins excreted by male and female rats during puberty were similar and of low molecular weight. In adulthood, moderate increases in diffuse staining of 26 and 21 kd occurred in female urine. In males the 17 kd protein increased dramatically and the 23 and 21 kd proteins moderately. The urine excretion of high-molecular-weight proteins (75 to 63 kd) increased with age in males (females not studied). Immunoblot studies with six sera showed allergens in urine of male and female rats of all ages, three of which were present in all urine (75, 68, and 21 kd). Three allergens (17, 16, and 15 kd) in female urine may be allergenically similar to the 17 kd allergen in adult male urine. Adult male urine allergens were studied further with sera from 83 rat-hypersensitive subjects. Major allergens were identified at 23, 21, and 17 kd, and all sera had IgE to one or more of these proteins. Twenty-seven percent had IgE to 68 and 63 kd allergens. Minor allergens were identified at 75, 51, and 44 kd.. Rat urine is an important source of the major allergens associated with rat hypersensitivity. Age and sex markedly influence the protein and allergenic constituents of rat urine.

Topics: Aging; Allergens; Animals; Electrophoresis, Polyacrylamide Gel; Female; Humans; Immunoblotting; Male; Proteinuria; Radioallergosorbent Test; Rats; Rats, Wistar; Sex Characteristics; Sodium Dodecyl Sulfate

Renal toxicity is a common manifestation to the exposure of laboratory animals and humans to a wide range of xenobiotics. Traditional methods for evaluating renal damage by clinical chemistry such as blood urea nitrogen (BUN) and serum creatinine are not sensitive to early, mild changes. The use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to measure the molecular weight spectrum of urinary proteins allows for an evaluation of the functional changes associated with renal damage. The ability of the kidney to filter and reabsorb proteins is related to the functional ability of glomeruli and the proximal tubules. Gentamicin sulfate produces injury to the S-1 and S-2 segments of the proximal tubule in laboratory animals and humans. While severe damage to the tubules is associated with increased BUN, serum creatinine, and N-acetyl-beta-glucosiminadase (NAG), mild injury is not detected by these means. The evaluation of urinary proteins by SDS-PAGE demonstrated renal toxicity at a dose of 6 mg/kg after 2 days of sc treatment. The NAG: creatinine ratio was shown to be elevated after 2 days of treatment at 63 mg/kg. The use of SDS-PAGE as described in this paper provides a sensitive method for detecting renal injury.

Topics: Acetylglucosaminidase; Animals; Blood Urea Nitrogen; Creatinine; Dose-Response Relationship, Drug; Electrophoresis, Polyacrylamide Gel; Female; Gentamicins; Kidney; Kidney Diseases; Kidney Tubules, Proximal; Male; Molecular Weight; Proteinuria; Rats; Rats, Sprague-Dawley; Sodium Dodecyl Sulfate

Of sixty patients with IgA nephritis, none had CRF at first examination, 13 developed CRF with creatinine above 1.6 mg/dl within 6 years. Among these patients who had analysis of proteinuria by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), 31 patients had middle molecular weight (MMW) proteinuria alone (pattern 1), 10 had MMW and Low MW (LMW) or tubular proteinuria (pattern 2), 10 had high MW (HMW) and MMW proteinuria (Pattern 3) and 9 had HMW, MMW and LMW proteinuria (Pattern 4). At the end of a follow up period of 6 years (1983-1989) patients with mixed proteinuria had a higher incidence of chronic renal failure (CRF), 11/29 (38%) compared to those with pattern 1 proteinuria, 2/31 (6%) (chi 2 = 8.7, p less than 0.005). Based on the glomerular selectivity index (GSI), 19 patients had nonselective proteinuria but they did not have a higher incidence of CRF. By the selectivity index (SI), 18 patients had nonselective proteinuria and they showed a significantly higher incidence of CRF. Compared to the 41 patients who did not have LMW proteinuria, 19 patients with LMW proteinuria had more severe proteinuria. After a follow-up period of 6 years, patients with LMW proteinuria had a higher incidence of CRF (10% versus 47%, p less than 0.001). The presence of LMW proteinuria indicates a less favourable outcome and the pattern of proteinuria as assessed by the SDS-PAGE appears to be a better prognostic index in IgA nephritis than the SI and the GSI.

Topics: Adult; Electrophoresis, Polyacrylamide Gel; Female; Follow-Up Studies; Glomerulonephritis, IGA; Humans; Incidence; Kidney Failure, Chronic; Male; Molecular Weight; Proteinuria; Sodium Dodecyl Sulfate

Topics: Electrophoresis, Polyacrylamide Gel; Humans; Molecular Weight; Proteins; Proteinuria; Sodium Dodecyl Sulfate; Urinary Calculi

The recent introduction of the PhastSystem, an automatic electrophoresis and staining system with precast gradient-gels, allows rapid and reproducible analysis of proteinuria in patients suffering from renal injury. A routine method for sodium dodecyl sulfate-polyacrylamide gradient gel electrophoresis (SDS-PAGE) and silver staining of unconcentrated urine specimens in the PhastSystem is described and compared to our conventional "macro"-method with self-cast SDS-polyacrylamide gradient gels. The method described for the PhastSystem using 0.3 microL sample volumes and an 8-25% polyacrylamide gradient gel leads to highly reproducible results within 1.5 h. Before electrophoresis urine specimens were neither concentrated nor dialyzed. Samples with a protein concentration exceeding 5 mg/mL had to be diluted 1:5 (v/v). Analysis and documentation of PhastGels appeared as easy as with our conventional SDS-PAGE. Protein bands could reliably be identified by Western blotting. Urine and serum proteins, separated in PhastGels, were electrophoretically transferred to nitrocellulose and detected with specific antibodies against human albumin, transferrin, alpha-1-antitrypsin and IgG. Comparison of several standard kits for molecular weight determination revealed considerable differences concerning the quality of protein separation patterns. Availability of precast gels and automatization of SDS-PAGE and staining allows easy standardization of urine SDS-PAGE among clinical routine laboratories.

Topics: Blood Proteins; Blotting, Western; Electrophoresis, Polyacrylamide Gel; Humans; Proteinuria; Silver; Sodium Dodecyl Sulfate; Staining and Labeling

The analysis of urine obtained from fetuses with hydronephrosis, seen on ultrasound, can give a misleading assessment of residual renal function. Additional parameters for assessment of fetal renal function would be helpful. We have used SDS-polyacrylamide gel electrophoresis to separate urinary proteins from a fetus with obstructive uropathy and severe oligohydramnios, already present at 18 weeks of gestation. The dilated urinary bladder of the fetus was successfully shunted in utero with a double pigtail catheter which worked for 17 weeks, and a boy without renal or pulmonary insufficiency was born at 36 weeks. In this case the prenatal protein analysis by electrophoresis was a better indicator of the ultimate good pregnancy outcome than the evaluation of urinary electrolytes and osmolarity alone. We therefore suggest the addition of this test to the profile of renal function studies performed on fetal urine.

Topics: Electrophoresis, Polyacrylamide Gel; Female; Fetal Diseases; Humans; Hydronephrosis; Kidney; Pregnancy; Proteinuria; Sodium Dodecyl Sulfate; Urinary Catheterization

We adapted the pyrogallol red-molybdate method for total urinary protein to the Cobas Bio centrifugal analyzer. The method is simple, rapid, sensitive, and inexpensive. Addition of 25 mg of sodium dodecyl sulfate per liter to the reagent modifies protein reactivities so that the chromogenicity of human gamma globulins is the same as that of albumin. Results by this method and a comparison method that included gel filtration and a modified biuret reaction correlated well (r = 0.951).

Topics: Adult; Autoanalysis; Coloring Agents; Humans; Molybdenum; Proteinuria; Pyrogallol; Quality Control; Reference Values; Sodium Dodecyl Sulfate

Various Coomassie Blue reagents, containing either increased dye concentration or added sodium dodecylsulphate, were compared with a biuret method for the assay of total protein in urine. When immunoglobulin free light chain protein or immunoglobulin paraprotein were present, results from the Coomassie Blue methods were up to 50% lower than with the biuret method; increased dye concentration did not improve comparability substantially, but the addition of sodium dodecylsulphate reduced the bias to about 20%. When neither free light chain protein nor immunoglobulin paraprotein was present, results from the Coomassie Blue methods were only about 30% lower. The addition of sodium dodecylsulphate reduced this bias to 10%. Correlations between the biuret and the Coomassie Blue method were best when the Coomassie Blue reagent contained 40 mg/L sodium dodecylsulphate (r better than 0.98 in all groups; p less than 0.001).

Topics: Binding, Competitive; Coloring Agents; Humans; Proteinuria; Rosaniline Dyes; Sodium Dodecyl Sulfate

Proteins in normal human urine were clearly fractionated into 26 bands with molecular weights from 14,000 to 230,000 by means of one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) coupled with silver staining. The main band contained uromucoid, and the second main band had albumin. However, when urine samples from healthy persons were electrophoresed in the absence of SDS using polyacrylamide gel or agarose gel, or a cellulose acetate membrane, albumin but not uromucoid, frequently formed the main protein band. It is suggested that this is due to the complexing of uromucoid subunits to form a large molecule which cannot penetrate into the gel. In order to correctly fractionate all the proteins contained in normal human urine, it was concluded that it was best to treat a urine sample with SDS with pre-condensation, fractionate it by SDS-PAGE and stain fractionated proteins by a highly sensitive method such as silver staining.

Topics: Adult; Albuminuria; Electrophoresis, Agar Gel; Electrophoresis, Disc; Electrophoresis, Polyacrylamide Gel; Female; Humans; Male; Middle Aged; Proteinuria; Sodium Dodecyl Sulfate

We describe an investigation of proteinuria using Pharmacia PhastSystemTM electrophoresis apparatus. The analysis of urinary proteins by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) of unconcentrated urine followed by silver staining took about 2 h and could clearly demonstrate tubular dysfunction or glomerular damage in urines with a negative or only trace-positive dip-stick test for protein. In addition, we show the identification of urinary proteins by immunoblotting from SDS-PAGE gels and the characterisation of Bence-Jones proteins by isoelectric focusing (IEF) and immunoblotting.

Topics: Autoanalysis; Bence Jones Protein; Child; Diabetic Nephropathies; Electrophoresis, Polyacrylamide Gel; Female; Humans; Isoelectric Focusing; Lupus Erythematosus, Systemic; Male; Muscular Diseases; Proteinuria; Sodium Dodecyl Sulfate

The Coomassie Brilliant Blue G-250 method for urinary proteins underestimates urinary immunoglobulin light chains when albumin or pooled serum is used as the protein standard. The specific color yields of these and other proteins can be brought closer together by adding sodium dodecyl sulfate to the reagent; however, there is some loss of sensitivity. We found such a reagent to be satisfactory for assaying urinary proteins on studying 43 patients with light-chain proteinuria, 19 of whom had multiple myeloma and six possible multiple myeloma.

Topics: Adult; Aged; Aged, 80 and over; Coloring Agents; Female; Humans; Immunoglobulin kappa-Chains; Immunoglobulin lambda-Chains; Male; Middle Aged; Multiple Myeloma; Proteinuria; Rosaniline Dyes; Sodium Dodecyl Sulfate; Spectrophotometry

Immunoblotting methods have allowed the identification of 14 urinary proteins. From this study, it appears that some low-relative-molecular mass proteins are derived from disrupted high-relative-molecular mass proteins. The only reliable bands for the diagnosis of tubular lesions, on the polyacrylamide gel stained with Coomassie Blue, are retinol-binding-protein and beta 2-microglobulin. alpha 1-microglobulin can overlap with breakdown products of albumin. The visualization of Ig light chains and lysozyme is rather poor after Coomassie Blue stainings and the accurate identification of these bands may be only improved by the use of immunoblotting.

Topics: Collodion; Electrophoresis, Polyacrylamide Gel; Evaluation Studies as Topic; Humans; Immunochemistry; Immunoglobulin G; Kidney Diseases; Kidney Tubules; Orosomucoid; Proteinuria; Sodium Dodecyl Sulfate

Topics: Diabetes Mellitus, Type 2; Diabetic Nephropathies; Electrophoresis, Polyacrylamide Gel; Female; Humans; Male; Middle Aged; Proteinuria; Sodium Dodecyl Sulfate; Urinary Tract Infections

We applied a simple sodium dodecyl sulfate-polyacrylamide gel electrophoresis method to urine. The method, developed for serum protein analysis (Clin Chem 1984;30:475-9), has a high sample throughput and gives excellent resolution with unconcentrated urine. It clearly distinguishes and characterizes proteinuric urine (7.5 microL) by Coomassie Blue staining and gives complex silver-stained patterns with nonproteinuric urine (2 microL). The former is recommended for routine clinical screening, the latter for research purposes.

Topics: Electrophoresis, Polyacrylamide Gel; Humans; Proteinuria; Rosaniline Dyes; Sodium Dodecyl Sulfate; Urine

Immunoblots of several urinary low-molecular-mass proteins can be very useful in investigations of pathological proteinuria. However, use of certain commercial antisera in such procedures leads to artifacts corresponding to nonspecific bands; e.g., immunoglobulins from nonimmunized rabbit serum may bind to human urinary proteins, and this binding is not inhibited by Triton X-100. We have developed a procedure to improve the specificity of detection of urinary low-Mr proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, by immunoblotting with commercial antisera: we treat the protein blot with a mixture of mercaptoethanol and sodium dodecyl sulfate before incubation with the first antiserum. This allows direct use of commercial antisera without prior absorption of contaminating antibodies.

Topics: Alpha-Globulins; beta 2-Microglobulin; Electrophoresis, Polyacrylamide Gel; Humans; Immune Sera; Immunosorbent Techniques; Mercaptoethanol; Methods; Muramidase; Octoxynol; Polyethylene Glycols; Proteinuria; Sodium Dodecyl Sulfate

We described a rapid and sensitive method for urinary proteins disc-electrophoresis on unconcentrated urine, by using polyacrylamide-SDS gel in discontinuous gradient. Correlation with a conventional polyacrylamide agarose slab electrophoresis on concentrated urine is good, our technique being more sensitive when proteinuria is low. It had proved to be useful for routine detection of renal failures in clinical laboratories.

Topics: Acrylic Resins; Albuminuria; Alpha-Globulins; beta 2-Microglobulin; Electrophoresis, Disc; Humans; Molecular Weight; Myeloma Proteins; Proteinuria; Sodium Dodecyl Sulfate; Transferrin; Urine

The molecular weight analysis of urinary proteins can provide useful diagnostic information. For this purpose a routine method of sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-electrophoresis) is described. The main problem in the introduction of this method into the clinical laboratory lies in the availability of "ready-to-use" electrophoresis gels with good and reproducible quality. A device for gel production is therefore described, which is easily constructed from plastic package materials. Polyacrylamide gels are made batch-wise and in advance. They are suitable for various types and techniques of horizontal electrophoresis. Further, a method for analysis of urinary proteins is described, which permits the simultaneous analysis of 22 unconcentrated urine samples within a 3-hour-electrophoresis run. A specially optimized Coomassie blue staining method (overnight) ensures detection of proteins in the concentration range of milligrams per litre. Electrophoretic analyses were documented by photocopying the finished electrophoresis gels.

Topics: Electrophoresis, Polyacrylamide Gel; Humans; Kidney Transplantation; Proteinuria; Sodium Dodecyl Sulfate

It is suggested that Tamm-Horsfall protein, a specific renal glycoprotein, may be involved in the pathogenesis of some renal diseases. In cadmium nephropathy and Fanconi syndrome (primary tubular diseases of the kidney) an increased excretion rate of Tamm-Horsfall protein has been observed. Balkan endemic nephropathy is a chronic tubulointerstitial disease of unknown etiology, most probably a primary disease of the kidney tubules with secondary reaction of the interstitial tissue. Investigation of Tamm-Horsfall proteinuria in Balkan endemic nephropathy has shown that subjects living in the area where this condition is prevalent have a significantly higher Tamm-Horsfall protein /creatinine ratio than those living in the control area where the condition has not been observed. Differences in this ratio among diseased, suspect and subjects "at risk" were not observed, despite differences in their glomerular filtration rates. But excretion of Tamm-Horsfall protein per litre of glomerular filtrate was significantly different among diseased, suspect and subjects "at risk" and significantly higher compared to control subjects. a relatively significant correlation was obtained between Tamm-Horsfall protein excretion rate and glomerular filtration rate as measured by creatinine clearance in both control and subjects living in the area of Balkan endemic nephropathy. Determination of Tamm-Horsfall protein in urine together with determination of proteinuria by electrophoresis on cellulose acetate membranes as a screening procedure, and by SDS -electrophoresis in polyacrylamide gell may be useful laboratory tests in detecting this nephropathy.

Topics: Adolescent; Adult; Aged; Balkan Nephropathy; Creatinine; Electrophoresis, Cellulose Acetate; Electrophoresis, Polyacrylamide Gel; Female; Humans; Male; Middle Aged; Mucoproteins; Nephritis, Interstitial; Proteinuria; Sodium Dodecyl Sulfate; Uromodulin

The pattern of proteinuria found in patients during the administration of methotrexate (MTX) or aminoglycosides (AG) and in cadmium or Balkan nephropathy was investigated using the technique of sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). As renal tubular dysfunction increased, measured by the urinary concentration of 4 low molecular mass (LMr) proteins, SDS-PAGE bands appeared in the following order of molecular mass (Mr): 59, 44, 31, then below 31 000. The presence of bands less than 31 000 was not an early indicator of drug-induced renal damage. Tubular proteinuria could be monitored more easily by the serum and urinary measurement of any one of the LMr proteins: alpha-1-microglobulin (alpha 1m), retinol-binding protein (RBP), beta-2-microglobulin (beta 2m), except alpha-1-acid glycoprotein (AGP), than by SDS-PAGE.

Topics: Adult; Aged; Aminoglycosides; Balkan Nephropathy; Cadmium Poisoning; Creatinine; Electrophoresis, Polyacrylamide Gel; Female; Humans; Kidney Tubules; Male; Methotrexate; Middle Aged; Molecular Weight; Nephritis, Interstitial; Proteinuria; Sodium Dodecyl Sulfate

Topics: Biuret Reaction; Humans; Indicators and Reagents; Methods; Proteinuria; Rosaniline Dyes; Sodium Dodecyl Sulfate; Ultrafiltration

Topics: Collodion; Electrophoresis, Polyacrylamide Gel; Filtration; Humans; Molecular Weight; Proteinuria; Sodium Dodecyl Sulfate

Topics: Chemical Phenomena; Chemistry; Chromatography, Gel; Humans; Indicators and Reagents; Proteinuria; Rosaniline Dyes; Sodium Dodecyl Sulfate

Patterns of urinary proteins were examined on sodium dodecyl sulphate polyacrylamide gels and compared to those obtained using high voltage agarose electrophoresis. Both were found to be well adapted to routine laboratory analysis although the former had more sensitivity for early renal changes. Glomerular, mixed and tubular patterns were identifiable, and special precautions and pitfalls whilst interpreting the latter were pointed out. Restricted alpha 1 antitrypsin was shown in renal transplant proteinuria and its possible mechanisms were discussed. Molecular masses and frequencies of several discrete bands in heavy proteinurics were calculated, and attempts to identify them were made with reference to standard proteins.

Topics: Densitometry; Electrophoresis, Disc; Humans; Kidney Glomerulus; Kidney Tubules; Molecular Weight; Proteinuria; Sodium Dodecyl Sulfate

Topics: Electrophoresis, Disc; Humans; Proteins; Proteinuria; Sodium Dodecyl Sulfate

We describe a procedure for the convenient separation of proteins by sodium dodecyl sulfate/polyacrylamide gel electrophoresis of urine from cases of renal disease. A precipitation method that requires no special apparatus was used to concentrate the urinary proteins; for electrophoretic separation we used a commercially supplied polyacrylamide/cellulose gel slab. This method seems to be valuable for investigation of proteinuria; we recommend it for routine use.

Topics: Chemical Precipitation; Electrophoresis, Polyacrylamide Gel; Humans; Iodoacetamide; Proteinuria; Sodium Dodecyl Sulfate; Ultrafiltration

Topics: Adult; Aged; Creatinine; Diabetic Nephropathies; Electrophoresis, Polyacrylamide Gel; Female; Humans; Male; Middle Aged; Proteinuria; Sodium Dodecyl Sulfate

Qualitative analysis of urinary proteins is contrasted with histological findings of 45 renal biopsies performed in patients with chronic glomerulonephritis. Compared to electrophoresis on cellulose acetate and immunoelectrophoresis, a method using polyacrylamide gel after sodium dodecylsulfate treatment makes for more refined and objective differentiation of protein abnormalities. On the whole, proteinuria of the selective glomerular or physiological type predominates in the event of minimal change or membranous lesions. The non-selective type is found more frequently with diffuse proliferative or membranoproliferative glomerulonephritis (p less than 0.025). There are, however, too many exceptions to this rule to allow certainty, and a precise diagnosis of the particular type of glomerulonephritis is thus only possible histologically. Each type of histological involvement may cause almost any of the qualitative abnormalities of proteinuria. On the other hand, qualitative analysis of urinary proteins is useful for the detection of glomerulonephritis. A glomerular type of proteinuria may sometimes reveal involvement of kidneys at a time when, quantitatively, there is no proteinuria. In cases of orthostatic proteinuria a persistent glomerular type of tracing in recumbency suggests an organic kidney ailment. All patients in this series had a glomerular type of proteinuria when excretion was pathological, thus allowing a distinction from pure tubular involvement. 10 patients of the group, however, although they clearly had glomerular lesions (3 were diffuse proliferative glomerulonephritis) showed perfectly normal proteinuria both quantitatively and qualitatively. This was the case in systemic lupus erythematosus where kidney biopsy was performed without clinical suspicion of renal involvement. In summary, qualitative abnormalities of proteinuria call attention to underlying glomerulonephritis, although no distinction can be made between the various forms and there may be no detectable abnormality even in the event of major kidney involvement.

Topics: Adolescent; Adult; Aged; Electrophoresis, Polyacrylamide Gel; Female; Glomerulonephritis; Humans; Kidney Glomerulus; Lupus Erythematosus, Systemic; Male; Middle Aged; Proteinuria; Sodium Dodecyl Sulfate

Topics: Electrophoresis, Polyacrylamide Gel; Humans; Kidney Glomerulus; Male; Middle Aged; Molecular Weight; Nephritis; Proteinuria; Sodium Dodecyl Sulfate

The authors present a variant of the technique of sodium dodecylsulfate acrylamide gel electrophoresis (SDS-PAA) reported by Weber and Osborn, for the analysis of urinary proteins. SDS-PAA separates the proteins chiefly according to their molecular radius. SDS-PAA, as compared to acetate cellulose electrophoresis and immunoelectrophoresis, gives better resolution and may be recommended for the investigation of proteinuria.

Topics: Electrophoresis, Cellulose Acetate; Electrophoresis, Polyacrylamide Gel; Evaluation Studies as Topic; Humans; Immunoelectrophoresis; Proteins; Proteinuria; Sodium Dodecyl Sulfate

Topics: Adolescent; Adult; Aged; Child; Diabetes Mellitus; Electrophoresis, Polyacrylamide Gel; Female; Humans; Hypertension; Kidney Diseases; Kidney Glomerulus; Kidney Tubules; Male; Methods; Middle Aged; Molecular Weight; Nephrotic Syndrome; Proteinuria; Sodium Dodecyl Sulfate; Urine

Topics: Dansyl Compounds; Electrophoresis, Disc; Electrophoresis, Polyacrylamide Gel; Molecular Weight; Proteinuria; Sodium Dodecyl Sulfate

Topics: Animals; Cellulose; Chromatography, Gel; Diabetes Mellitus; Dialysis; Electrophoresis; Electrophoresis, Polyacrylamide Gel; Evaluation Studies as Topic; Horses; Humans; Hydrogen-Ion Concentration; Hypertension; Immunoelectrophoresis; Kidney Diseases; Methods; Molecular Weight; Proteinuria; Sodium Dodecyl Sulfate; Time Factors; Ultrafiltration

Topics: Albuminuria; Bence Jones Protein; Blood Proteins; Chromatography, Gel; Electrophoresis, Disc; Fructose-Bisphosphate Aldolase; gamma-Globulins; Humans; Kidney Diseases; Kidney Glomerulus; Kidney Tubules; L-Lactate Dehydrogenase; Methods; Molecular Weight; Muramidase; Ovalbumin; Pepsin A; Protein Binding; Proteinuria; Sodium Dodecyl Sulfate

Topics: Acute Kidney Injury; Blood Proteins; Creatinine; Electrophoresis, Polyacrylamide Gel; Fanconi Syndrome; Glomerular Filtration Rate; Humans; Immunoglobulin Fragments; Immunoglobulin G; Kidney; Kidney Diseases; Kidney Glomerulus; Nephrons; Nephrotic Syndrome; Proteinuria; Renal Tubular Transport, Inborn Errors; Sodium Dodecyl Sulfate; Uremia