242
TAKEHIKO KOJI
~ 1·
• • . .
. .,
,oJ.
,
.
' ..
••
Fig. 4. In situ detection of CRE-binding proteins by Southwestern histochemistry in the fresh frozen sections of rat small intestine (Koji et al. 1994). The fresh
frozen sections of rat small intestine were fIxed with 4% paraformaldehyde in
PBS and reacted with T-T dimerized CRE double-stranded oligo-DNA probe
(CRE); the left panel is a low-power view and the right panel is a high-power
view. (Non probe); a section was processed omitting only the probe DNA.
(Comp); a section was reacted with the CRE probe in the presence of an excess
amount of non-labeled CRE DNA (102-fold). In CRE panels, the strong staining
of epithelial nuclei of small intestine was found, whereas in aNon probe section
and a Comp section no signal was detected.
Before use, the double-stranded eRE oligo-DNA was T-T dimerized by UV irradiation. When the sections of those tissues,
which were known to be responsive to cAMP, were reacted with
the haptenized probe, the brain and intestinal epithelial nuclei
were heavily stained and the staining intensities could be altered
by physiological manipulation (Koji et al. 1994).
Although the list of transcription factors and their responsive
elements is rapidly growing, antibodies are not available for
most of these newly identified proteins. This method should
TAKEHIKO KOJI
~ 1·
• • . .
. .,
,oJ.
,
.
' ..
••
Fig. 4. In situ detection of CRE-binding proteins by Southwestern histochemistry in the fresh frozen sections of rat small intestine (Koji et al. 1994). The fresh
frozen sections of rat small intestine were fIxed with 4% paraformaldehyde in
PBS and reacted with T-T dimerized CRE double-stranded oligo-DNA probe
(CRE); the left panel is a low-power view and the right panel is a high-power
view. (Non probe); a section was processed omitting only the probe DNA.
(Comp); a section was reacted with the CRE probe in the presence of an excess
amount of non-labeled CRE DNA (102-fold). In CRE panels, the strong staining
of epithelial nuclei of small intestine was found, whereas in aNon probe section
and a Comp section no signal was detected.
Before use, the double-stranded eRE oligo-DNA was T-T dimerized by UV irradiation. When the sections of those tissues,
which were known to be responsive to cAMP, were reacted with
the haptenized probe, the brain and intestinal epithelial nuclei
were heavily stained and the staining intensities could be altered
by physiological manipulation (Koji et al. 1994).
Although the list of transcription factors and their responsive
elements is rapidly growing, antibodies are not available for
most of these newly identified proteins. This method should