sodium-dodecyl-sulfate and pyrogallol-sulfonphthalein

We have investigated the effect of sodium dodecyl sulfate (SDS) upon the response of the Coomassie Brilliant Blue (CBB) and Pyrogallol Red-molybdate (PRM) protein dye-binding assays to interference from aminoglycosides, ampholytes, detergents, phenothiazines, reducing agents, and miscellaneous substances previously reported to interfere with the assays. The CBB assay was less prone to interference than the PRM assay but gave positive interference with the detergents and the phenothiazines and negative interference with dextran sulfate. The PRM assay gave positive interference with the aminoglycosides, ampholytes, and phenothiazines and negative interference with SDS, citric acid, dextran sulfate, EDTA, oxalic acid, and tartaric acid. The level of interference varied in the presence of different proteins (albumin, gamma globulin, alpha1-acid glycoprotein, or lysozyme) and increased when SDS was added to the dye reagents.

Topics: Indicators and Reagents; Pyrogallol; Rosaniline Dyes; 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

The Pyrogallol Red Molybdate (PRM) and Coomassie Brilliant Blue (CBB) protein dye-binding assays have been applied to samples of cerebrospinal fluid (CSF) to investigate protein concentration by dye precipitation prior to sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The protein concentration values of the CSF samples (N=62) showed good agreement between the PRM and CBB assays as indicated by linear regression analysis (y(PRM)=1.033x(CBB)+1.004 in units of mg/l, r=0.99) but the PRM assay was optimal for protein concentration as the PRM protein-dye complex was less soluble allowing protein recovery over a wider working range. Dye precipitation using PRM is recommended as a simple, rapid and economic method for protein concentration of samples of CSF prior to SDS-PAGE.

Topics: Cerebrospinal Fluid Proteins; Chemical Precipitation; Coloring Agents; Electrophoresis, Polyacrylamide Gel; Humans; Nervous System Diseases; Pyrogallol; Regression Analysis; Rosaniline Dyes; Sodium Dodecyl Sulfate; Solubility

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