 Apoptosis Inducers |
All three are known to induce apoptosis. Valinomycin is used
to induce mitochondrial potential disruption. Etoposide is
a DNA synthesis inhibitor that induces double stranded and
single stranded DNA breaks. Staurosporine is a phospholipid/calcium-dependent
protein kinase inhibitor that prevents ATP binding.
|
|
| Apoptosis Grade™ PBS Buffer and Water |
The 10X PBS buffer is a high quality preparation manufactured
without magnesium or cal-cium, and is suitable for cell resuspension
and washing in the in situ apoptosis detection procedure,
CometAssay™ (Cat# 4250-50-K) applications, and a variety
of other applica-tions requiring ultrapure PBS buffer.
The water is purified through a reverse osmosis system and
multistage nanoparticle filtration system assuring greater
than 18 megohm-cm2 resistivity is provided in a convenient
500 ml size. The water is then sterilized using a unique autoclaving
process to assure the highest quality.
|
|
| Controls |
Trevigen offers several different types of control slides
to develop familiarity with the methods of in situ apoptosis
labeling and to determine if reagents are working optimally.
The Cell Culture Control Slides are useful for researchers
studying apoptosis in cell culture sys-tems, whereas the Tissue
Control Slides are helpful when investigating tissue specimens.
EpiDerm™ Control slides (Cat# 4800-30-42) are sections of synthetic
human skin tailored for use with DermaTACS™.
TACS-Nuclease is a novel DNA endonuclease that is used to
prepare a positive control from your own samples. The nuclease
can simply be added to the labeling reaction mixture or if
preferred in a separate step. DNA fragments formed by the
action of this enzyme serve as substrates for labeling, producing
a positive signal in virtually every cell.
|
|
| Counterstains |
|
Trevigen offers a variety of counterstains for use in our
TUNEL assay kits. Blue
counterstain unlike Methyl Green works well in neuronal tissue,
and is also a good choice for double-labeling with brown or
red detection systems. Nuclear Fast Red is the ideal counterstain
for Trevigen kits employing the TACS® Blue Label chromogenic
substrate. Red Counterstain C provides an alternative for
a red counterstain. Our Methyl Green stain is free of cresyl
violet contamination and provides excellent contrast with
DAB detection systems.
|
|
| Mounting Media |
|
Trevigen Mounting Medium is a high quality, optically clear,
toluene based mounting medium. This mounting medium is ideal
for mounting samples processed with our TACS® In Situ
Apoptosis Detection kits using either diaminobenzidine or
TACS® Blue Label™. Mounting samples helps protect the
specimen from damage and helps produce a clear image for microscopic
visualization and photography. For fluorescent samples, use
Trevigen Fluorescence Mounting Medium (Cat# 4866-20).
Trevigen Fluorescence Mounting Medium is specially designed
for use with our In Situ Apoptosis Detection kits that use
fluorescein as the detection method. The mounting medium hardens
completely in a short time, preserving the sample and providing
an air-tight seal under the glass coverslips.
|
|
| Permeabilization
Reagents for In Situ Apoptosis Detection |
|
Proteinase K is used for permeabilizing cells and tissues
prior to labeling using any of the TACS® In Situ Apoptosis
Detection Kits.
NeuroPore is a non-proteolytic permeabilization and blocking
reagent developed for use with the NeuroTACS® In Situ
Apoptosis Detection kit (Cat# 4820-30-K). Many tissues of
the central nervous system are fragile and do not tolerate
harsh proteolytic treatment. NeuroPore provides a gentle method
for the permeabilization of tissues prior to in situ labeling.
In addition, NeuroPore is an ideal antibody diluent for use
in double labeling experiments: antigenic determinants are
retained during permeabilization and can be detected using
standard immunohistochemical techniques in conjunction with
in situ detection of apoptosis.
Cytonin offers an alternative to Proteinase K for permeabilizing
cells and tissues prior to labeling using any of the TACS®
In Situ Apoptosis Detection kits. Cytonin is a protease free,
saponin-based buffer designed specifically for in situ detection
of apoptosis. Cytonin should be used when protease treatment
must be avoided. For example, it may be used for tissues with
low cellularity or little connective tissue that do not withstand
protease treatment. Cytonin IHC (see below) is recommended
for double labeling experiments for the detection of DNA fragmentation
in conjunction with immunohistochemistry on antigens that
are sensitive to proteolysis.
Cytonin-IHC a non-proteolytic, saponin-based permeabilization
and blocking reagent designed specifically for double labeling
experiments using TACS® In Situ Apoptosis Detection kits, and
a primary antibody of your choice. Cytonin IHC is used as
a convenient time-saving antibody diluent for immunohistochemistry,
allowing you to simultaneously perform your primary antibody incubation and permeabilization
of your sample for in situ detection of apoptosis. In fact,
use Cytonin IHC as a diluent for your secondary antibody too.
|
|
| Slides and Coverslips |
|
These slides are specially treated to
promote electrostatic adhesion of your tissue or cell samples.
The slides are frosted on one end allowing you to clearly
identify each sample by labeling with a pencil or histology
pen. The slides are precleaned and ready for use.
These slides have the same coating as our treated glass microscope
slides (Cat# 4861-100) with an added 3 ring hydrophobic barrier
allowing you to process 3 different samples easily on one
slide. The barrier promotes better sample coverage and conserves
reagents. Also, the barrier makes the task of drying suspension
cells onto slides as easy as pipetting.
These coverslips are made of a specially treated rigid plastic
that produces a hydrophobic surface that assures rapid and
even distribution of reagents over the entire sample being
labeled. The coverslips should be used to conserve reagents
when larger sample areas are being labeled. Unlike glass coverslips
that may not wet evenly and may be too heavy to allow sufficient
reagent to contact the sample, these coverslips float easily
on small volumes of labeling reagents. Unlike wax film or
other commercially available coverslips, these coverslips
are rigid, preventing uneven distribution of labeling solutions.
The coverslips are packaged in rows of 5 with a protective
film coating.
These coverslips are optically clear glass ideal for mounting
onto your specimens prior to viewing. The coverslips are clean
and ready for use. The 22 x 60 mm size assures complete coverage
when using our 3 ring barrier slides (Cat# 4864-100), or when
large samples are being processed. These may be used with
Trevigen Mounting Medium (Cat# 4865-25), or Trevigen Fluorescence
Mounting Medium (Cat# 4866-20).
|
|
| Streptavidin
Conjugates |
|
Trevigen offers a number of streptavidin conjugates to allow
flexibility in both in situ labeling, and our flow cytometric
based assays. Optimal concentrations range from 1:50 dilutions
to 1:1000 dilutions, depending upon application. The table
below summarizes the characteristics of each conjugate.
|
| |
Abs.
Max |
Em.
Max |
|
| Streptavidin-FITC |
492 nm
|
520 nm
|
"Green" |
| Streptavidin-AMCA |
345 nm |
445 nm
|
"Blue" |
|
|
| Strep-HRP |
|
Streptavidin, Mr 60,000, is covalently linked to horseradish peroxidase by a modification of the periodate method with a molar enzyme/protein ratio of 2.5:1. This conjugate binds four molecules of biotin with high affinity (Kd = 10-15 M-1), and is provided in liquid form with a proprietary stabilizer added, without preservative.
|
