Elsevier

Analytical Biochemistry

Volume 240, Issue 2, 5 September 1996, Pages 306-308
Analytical Biochemistry

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Comparison of Three Chemiluminescent Horseradish Peroxidase Substrates for Immunoblotting

https://doi.org/10.1006/abio.1996.0364Get rights and content

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    For sodium dodecylsulfate gel electrophoresis, 150 μg protein was placed per lane and Western blot was performed as previously described [21]. For antigen detection, a monoclonal C-MYC antibody (1:500, DAKO, Glostrup, Denmark) was used and visualized by a corresponding POD-marked secondary antibody (Dianova, Dörentrup, Germany) followed by the lumi-light method according to Mattson and Bellehumeur [22]. For immunohistochemical staining, 6-μm juvenile angiofibroma sections were deparaffinized.

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    Horseradish peroxidase-conjugated anti-rabbit IgG and anti-mouse IgG were used as secondary antibodies for the His and T7 antibodies, respectively. Immunocomplexes were revealed with an ECL kit according to the manufacturer's protocol (30). SDS-PAGE (12%) was carried out according to the method of Laemmli (31).

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