Immunodetection of nitrocellulose-adhesive proteins at the nanogram level after trinitrophenyl modification
✍ Scribed by J.Michael Wolff; Joachim Pfeifle; Michael Hollman; F.Alfred Anderer
- Publisher
- Elsevier Science
- Year
- 1985
- Tongue
- English
- Weight
- 772 KB
- Volume
- 147
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
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.