VectorCatalog2012.pdf? - Vector Laboratories

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Introduction
V E C T O R L A B O R A T O R I E S
With over 35 years of research, development, and
manufacturing experience Vector Laboratories has
acquired considerable expertise in the production of
immunofluorescence reagents. Our extensive range of
fluorescent reagents accommodates a variety of experimental
designs and levels of signal amplification.
Fluorochrome-Labeled Secondary Antibodies. Our affinity
purified antibodies are unmatched in quality for immunological
techniques. All antibodies are prepared using proprietary
immunization schedules that produce high affinity antibodies.
The antibodies are purified by affinity chromatography, and
cross-reactivities that are likely to interfere with specific labeling
are removed by solid phase adsorption techniques. Antibodies
are conjugated to ensure the optimal degree of labeling while
not compromising the specificity or affinity of the antibody.
See Figure 1.
Fluorochrome-Labeled Streptavidin and Avidin
Systems. Fluorescent signals can be amplified using our
biotinylated secondary antibodies followed by our highly
purified fluorescent streptavidin and avidin conjugates. These
fluorescent conjugates possess very low non-specific binding
properties and extremely high affinity for biotin. Using a
biotin/avidin or biotin/streptavidin detection system results in
an additional layer of amplification over a directly conjugated
secondary antibody. See Figure 2.
Amplification of Fluorescent Signal using Biotinylated
Anti-Streptavidin or Biotinylated Anti-Avidin. Biotinylated
Anti-Streptavidin and Biotinylated Anti-Avidin antibodies
provide an ideal method to further increase sensitivity.
These antibodies bind to streptavidin or avidin, respectively,
through both their antigen binding sites and also through
the covalently attached biotin residues. After the first
application of a fluorochrome-labeled streptavidin or avidin,
the signal is amplified by incubation with Biotinylated Anti-
Streptavidin or a Biotinylated Anti-Avidin antibody followed by
a second incubation with fluorochrome-labeled streptavidin
or avidin. This procedure results in the introduction of more
fluorochromes at the target site. See Figure 3.
w w w . V E C T O R L A B S . C O m
The figures below illustrate different experimental setups used
to achieve increasing levels of sensitivity.
Biotinylated
Secondary
Antibody
Biotinylated
Secondary
Antibody
Fluorescent Secondary
Antibody
Primary Antibody
Antigen
Fluorescent
(Strept)avidin
Figure 1
Figure 2
Second Layer of
Fluorescent
Strept(avidin)
Biotinylated
Anti-Streptavidin
or Anti-Avidin
First Layer of
Fluorescent
Strept(avidin)
Figure 3