This service
includes sectioning,
TUNEL labeling,
coverslipping
and labeling of
slides. As a result,
you will receive
up to 40 TUNEL-labeled
sections per brain
or per tissue
block ready for
microscopic observations.
To express our
appreciation,
a set of Nissl
(or H & E)
stained sections
adjacent to those
used for TUNEL-labeled
sections will
also be provided
for you without
additional cost.
Procedure:
Unfixed
frozen tissue
will be cut on
a cryostat and
mounted on proteinase-resistant
microscope slides
(see Products).
Sections cut through
various levels
(or the levels
of your choice)
will then be processed
on slides for
the detection
of neurons undergoing
apoptosis with
in situ DNA nick
end-labeling1.
This technique,
originally described
by Gavrieli et
al.(1992)1,
labels the nuclei
of neurons undergoing
DNA fragmentation.
This method is
based on the ability
of terminal deoxynucleotidyl
transferase to
catalyze incorporation
of biotinylated
deoxyuridines
onto the free
3'-hydroxyl termini
of DNA fragments,
which are considered
as one of the
most characteristic
features of apoptosis2,
3. The integrated
biotins are amplified
and visualized
by the avidin-biotin-complex
(ABC) method4
(see
NDT102).
Remarks:
· A quotation
is required before
placing an order.
· The investigator
needs to provide
unfixed frozen
tissue.
· Please contact
us for more information.
References:
1. Gavrieli Y,
Sherman Y, and
Ben-Sasson SA:
Identification
of programmed
cell death in
situ via specific
labeling of nuclear
DNA fragmentation.
J. Cell Biol.
119: 493-501,
1992.
2. Wyllie A. H.
(1980) Glucocorticoid-induced
thymocyte apoptosis
is associated
with endogenous
endonuclease activation.
Nature 284: 555-556.
3. Arends M. J.,
Morris R. G. and
Wyllie A. H. (1990)
Apoptosis: the
role of the endonuclease.
Amer. J. Pathol.
136: 593-608.
4. Hsu S. M.,
Raine L. and Fanger
H. (1981) Use
of avidin-biotin-peroxidase
complex (ABC)
in immunoperoxidase
techniques: a
comparison between
ABC and unlabeled
antibody (PAP)
procedures. J.
Histochem. Cytochem.
29: 577-580.
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