Western-blot identification of epitope.

<p>The 16-AA polypeptide of NS1-27 reacted with mAb 3G2 in Western-blot assay. Lane M, PageRuler Prestained Protein Ladder (Fermentas, Canada); Lane 1, GST-tag didn’t react with mAb 3G2; Lane 2, The band of NS1-27-GST fusion protein was visualized with mAb 3G2.</p

Sequences of the overlapping polypeptides from TMUV NS1 (SDSG strain, Accession number: KJ740747.1).

<p>Sequences of the overlapping polypeptides from TMUV NS1 (SDSG strain, Accession number: KJ740747.1).</p

Sequences of the truncated NS1-27 polypepide.

<p>Sequences of the truncated NS1-27 polypepide.</p

Primary screening of epitope with mAb 3G2.

<p>One 16-AA polypeptide of TMUV NS1 protein (NS1-27) was screened with mAb 3G2 by indirect ELISA. Mouse serum against TMUV NS1 protein and normal mouse serum were used as positive and negative controls, respectively. Each sample was detected in triplicate. Error bars were expressed as standard deviation of the means (n = 3). The mean value was statistically significant, calculated by the two-tailed Student’s unpaired t-test (*P < 0.05).</p

The accurate mapping of one B cell epitope with mAb.

<p>NS1-27 polypeptide was truncated from the carboxy and amino terminals. After the truncated peptides were expressed as a GST fusion protein, they were probed with mAb 3G2 by indirect ELISA respectively. The minimal unit of the peptide was the sequence of 8 AA and 3 AA truncated from the carboxy and amino terminals of NS1-27.</p

IFA and western-blot identification of mAb 3G2.

<p>A: Western-blot identification Lane 1, Control, GST-tag didn’t react with mAb 3G2; Lane 2, The band of NS1-GST fusion protein was reacted with mAb 3G2; Lane M, Blue plusIIprotein Marker (14-120kda, Transgen Biotech). B: IFA identification Monoclonal antibody against TMUV NS1 protein was used to perform IFA on TMUV-infected BHK-21 cells. BHK-21 cells infected with TMUV yielded significant fluorescence with six MAbs in the cytoplasm; Control BHK-21 cells didn’t yield any fluorescence.</p