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Clinical Chemistry 32: 811-815, 1986;
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Clinical Chemistry, Vol 32, 811-815, Copyright © 1986 by American Association for Clinical Chemistry

Reducing the artifacts produced by impure antisera in immunoblots of low-molecular-mass proteins in urine

JM Perini, B Dehon, T Marianne, A Klein and P Roussel

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.


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[Abstract] [Full Text] [PDF]


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Copyright © 1986 by the American Association for Clinical Chemistry.