Protein concentration of cerebrospinal fluid by precipitation with Pyrogallol Red prior to sodium dodecyl sulphate-polyacrylamide gel electrophoresis.
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.
Analytical Biochemistry Group, Institute of Pharmacy, Chemistry and Biomedical Sciences, School of Sciences, The University of Sunderland, SR1 3RG, Sunderland, UK. kath.williams@sunderland.ac.uk
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Williams
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Marshall
D002556:Cerebrospinal Fluid Proteins / Q000032:analysis*; D011232:Chemical Precipitation; D004396:Coloring Agents; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D009422:Nervous System Diseases / Q000134:cerebrospinal fluid / Q000175:diagnosis; D011748:Pyrogallol / Q000031:analogs & derivatives; D012044:Regression Analysis; D012394:Rosaniline Dyes; D012967:Sodium Dodecyl Sulfate; D012995:Solubility
D002556:Cerebrospinal Fluid Proteins; D004396:Coloring Agents; D012394:Rosaniline Dyes; D011748:Pyrogallol; C058731:pyrogallol sulfonphthalein; D012967:Sodium Dodecyl Sulfate; C004692:coomassie Brilliant Blue