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Figure 2 | Virology Journal

Figure 2

From: A simple, inexpensive, robust and sensitive dot-blot assay for equal detection of the nonstructural-1 glycoprotein of all dengue virus serotypes

Figure 2

Optimal quenching of endogenous human serum peroxidases in the DENV s/e NS1 glycoprotein dot-blot detection assay. Serial three-fold dilutions (90, 30 and 10 ng/ml) of immunoaffinity-purified DENV-2 s/e NS1 glycoprotein, and buffer without DENV-2 NS1 glycoprotein (0 ng/ml), prepared in undiluted human sera were added at 10 μl volumes to four sets of paired 5-mm squares marked on nitrocellulose membranes. These membranes were then dried and cut for reaction with each of the different solutions trialled for quenching endogenous human serum peroxidases (as shown), before being washed and blocked. These membranes were then processed by sequential reaction steps with MAb 2C4.6, peroxidase-labelled goat anti-mouse IgG (H&L) and 4-chloro-1-naphthol/3,3’diaminobenzidine (CND) substrate. The results were obtained from one of duplicate experiments performed that gave similar results.

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Virology Journal

ISSN: 1743-422X