Figure 1 | Cell Death & Disease

Figure 1

From: Potent antitumoral activity of TRAIL through generation of tumor-targeted single-chain fusion proteins

Figure 1

Biochemical analysis of scTRAIL and scFv–scTRAIL. (a) Scheme of the scTRAIL fusion proteins used in this study. L, leader peptide; scFv, human ErbB2-specific single-chain antibody fragment; F, FLAG tag; TRAIL, human TRAIL (aa 95–281). (b) Purified scTRAIL (lane 1) and scFv–scTRAIL (lane 2) were analyzed by SDS-PAGE (10%, reducing conditions) followed by immunoblot analysis (left, 250 ng protein/lane) or silver staining (right, 1 μg protein/lane). For immunoblotting, the monoclonal anti-FLAG M2 antibody and an alkaline phosphatase-conjugated secondary antibody was used. (c) The TRAIL variants were separated by gel filtration on a BioSuite250 column. Dotted lines indicate the retention time of the molecular weight standards apoferritin (443 kDa), β-amylase (200 kDa), bovine serum albumin (66 kDa) and carbonic anhydrase (29 kDa). Collected fractions were analyzed by immunoblotting as in b. For scTRAIL fractions 1–9 (every second fraction; first part), fractions 11–16 (second part) and fractions 19–26 (every second fraction; third part) were immunoblotted. For scFv–scTRAIL fractions 8–24 were immunoblotted

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