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A. -52C
B. w/o Tag
Fig. 4. Electron micrograph of VLP-52C. VLP-52C (A) and VLP with-
out tag (B) were observed under electron microscopy after negative staining
at a magnification of
×
60,000. Inserted bar indicates 100 nm.
Ab
none
-Tag
-HEV Cont.
+
-
+
-
++
-
-
Tag
Fig. 5. Surface exposure of the tag epitope on VLP-52C. Surface exposure of
the tag epitope on intact VLP-52C was examined by immunoprecipitation with
the anti-tag antibody. Antibodies used are indicated at the top of panel. None,
negative control without antibody;
-HEV, anti-
HEV antibody; cont., purified VLP-52C and VLP without tag were run as con-
trols. The second row indicates either VLP with (
α
-Tag, anti-tag antibody;
α
+
) or without the tag (
).
VLP without the tag (Fig. 4). Using two methods, we confirmed
that the inserted epitope tag was exposed on the surface. The intact
VLP-52C was immunoprecipitated with the anti-tag antibody, while
the anti-HEV antibody immunoprecipitated both VLP-52C and the
VLP without the tag (Fig. 5). Furthermore, the anti-tag antibody
specifically reacted with the intact VLP-52C in an ELISA (data not
shown). The results of immunoprecipitation and ELISA using intact
chimeric VLP suggest that the tag epitope is exposed on the surface
of the HEV-VLP.
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