| Autor: |
Shiohama, Yasuo, Tadasuke Naito, Matsuzaki, Toshio, Tanaka, Reiko, Tomoyose, Takeaki, Takashima, Hiroshi, Fukushima, Takuya, Yuetsu Tanaka, Mineki Saito |
| Rok vydání: |
2016 |
| DOI: |
10.6084/m9.figshare.c.3597737_d1.v1 |
| Popis: |
Additional file 1. Figure S1: Amino acid sequence of HBZ. A. The amino acid sequence of the HBZ protein is shown. The sequences of the peptides used to generate the anti-HBZ monoclonal antibodies are underlined. B. The histidine-tagged recombinant HBZ protein, which was produced by a wheat germ extract based cell free transcription and translation system (WEPRO7240H Expression kit; CellFree Sciences, Japan), was analyzed by SDS-PAGE. Gels were scanned and the levels of HBZ protein were quantified by BIO-RAD Quantity One 4.3.1 software using densities of known concentrations of BSA. Figure S2: Development of an ELISA for the detection of HBZ protein in HTLV-1-infected cells. HBZ protein levels in cell lysates were evaluated by in-house sandwich ELISA using mAbs against HBZ (i.e., clone #91-1 for capture and clone #20-H12 for detection). As shown, a 96-well flat-bottom plate was coated with an anti-HBZ mAb (clone #91-1: rat IgG1 mAb raised against peptide #3). Then, a second anti-HBZ mAb (clone #20-H12: mouse IgG1 mAb raised against peptide #2) conjugated to HRP was used as the detection antibody. Figure S3: Western blotting analysis of HBZ protein in HTLV-1-infected cell lines using various monoclonal antibodies (mAbs). HBZ protein expression in HTLV-1-infected cell lines was analyzed by western blotting using various anti-HBZ mAbs (i.e. P6-A7, #20-H12, #7-1, #91-1). Clone #91-1 (raised against peptide #3, rat IgG1) showed the highest sensitivity for detecting HBZ protein. Histone H3 was used as a loading control for the nuclear fraction. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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