Immunodetection of nitrocellulose-adhesive proteins at the nanogram level after 
trinitrophenyl modification

Wolff, JM; Pfeifle, J; Hollmann, M; Anderer, FA

doi:10.1016/0003-2697(85)90288-x

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Immunodetection of nitrocellulose-adhesive proteins at the nanogram level after trinitrophenyl modification

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Wolff, JM
Anderer Group, Friedrich Miescher Laboratory, Max Planck Society;

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Pfeifle, J
Anderer Group, Friedrich Miescher Laboratory, Max Planck Society;

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Hollmann, M
Anderer Group, Friedrich Miescher Laboratory, Max Planck Society;

/persons/resource/persons282624

Anderer, FA
Anderer Group, Friedrich Miescher Laboratory, Max Planck Society;

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引用

Wolff, J., Pfeifle, J., Hollmann, M., & Anderer, F. (1985). Immunodetection of nitrocellulose-adhesive proteins at the nanogram level after trinitrophenyl modification. Analytical Biochemistry, 147(2), 396-400. doi:10.1016/0003-2697(85)90288-x.

引用: https://hdl.handle.net/21.11116/0000-000D-9959-3

要旨

A method for protein detection on nitrocellulose membranes based on modification with 2,4,6-trinitrobenzenesulfonic acid and reaction with anti-trinitrophenyl (TNP) serum as first antibody followed by peroxidase-conjugated second antibody is described. Protein quantities between 1 and 3 ng can be detected in the dot test. This method was used in a double immunodetection procedure after electrophoretic transfer of proteins localizing first a distinct antigen with its specific antiserum followed by visualization of the complete protein pattern on the same blot by the TNP/anti-TNP method as described above. As only water-soluble reagents are employed no shrinkage of the membrane occurs. Furthermore, the method can be used in a simultaneous immunodetection procedure visualizing the specific antigen together with TNP marker proteins using a mixture of the specific antiserum and the anti-TNP serum as first antibody.