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Figure 5 | Virology Journal

Figure 5

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

Figure 5

Detection sensitivities of MAb 2C4.6 against DENV s/e NS1 glycoproteins stored at different temperatures. Serial two-fold dilutions from 512 to 2 ng/ml of immunoaffinity-purified s/e NS1 glycoprotein of each DENV serotype (DENV-1 to DENV-4) or buffer only (0), prepared in undiluted human sera were added at 10 μl volumes to four paired 5-mm squares marked on a nitrocellulose membrane. After being dried and treated with 3% H2O2/H2O for 5 min to quench the endogenous human serum peroxidases, this membrane was washed and blocked. After being washed and dried, the membrane was cut into four equal pieces, sealed in plastic bags and stored either at ambient temperature (mean 28°C), 4°C, -20°C or -80°C for 1 month. 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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