quality reagents for:
VECTASTAIN ABC Systems
VECTASTAIN Elite ABC Video Tutorial
||Tissue staining using the VECTASTAIN Elite ABC system. For additional instructional videos please see the Tutorials page under the Support link.
The VECTASTAIN® ABC systems are extremely sensitive due to the form and number of active enzyme molecules associated
with the preformed Avidin/Biotinylated enzyme Complex.
This complex takes advantage of two important properties
of avidin: 1) an extraordinarily high affinity for biotin (over
one million times higher than antibody for most antigens),
and 2) four biotin-binding sites. These properties allow
macromolecular complexes (ABCs) to be formed by mixing
Avidin DH (Reagent A) with its paired biotinylated enzyme
(Reagent B) prior to use. The ABC, once formed, remains
stable for many hours after formation and can be used for
several days after preparation.
The VECTASTAIN® ABC Reagent can be used to detect any
molecule that is biotinylated. This property gives the ABC
method great versatility in the types of targets that can be
detected as well as the types of applications in which it can
be employed. Biotinylated primary antibodies, secondaries,
lectins, neuronal tracers, nucleic acids, and ligands can be
effectively visualized in applications such as:
Protein and nucleic acid blot detection
In situ hybridization detection
All applications benefit from the high sensitivity, low
background, reproducibility, and economy of the VECTASTAIN® ABC system.
Choosing a VECTASTAIN® ABC Kit
1) Choose an enzyme system:
The VECTASTAIN® ABC Kits are available with a choice of
three different enzyme systems.
• Peroxidase (ABC, Elite® ABC, RTU or Kits)
• Alkaline Phosphatase (ABC-AP)
• Glucose Oxidase (ABC-GO)
2) Choose the biotinylated secondary antibody:
To detect a biotinylated target, you will need only the ABC
Reagent contained in the Standard Kit, followed by an
To detect an unlabeled primary antibody, you will need a
biotinylated secondary antibody that binds to the primary
antibody species chosen, the ABC Reagent, and an appropriate
substrate. For example, to detect a primary antibody made
in rabbit, the appropriate choice is a VECTASTAIN® ABC Kit
designated “Rabbit IgG”.
For additional versatility and convenience, Vector Laboratories
offers secondary antibody products that recognize more than
one species of primary antibody. Our Universal Biotinylated
Anti-Mouse/Rabbit IgG secondary antibody (Cat. No. BA-1400)
is a cocktail of biotinylated anti-mouse IgG and anti-rabbit IgG,
designed for use with both rabbit and mouse primary
antibodies. This antibody should not be used to stain tissues
from rodents or rabbits due to its reactivity with IgG present in
the tissues of these species. The Universal Pan-Specific Anti-
Mouse/Rabbit/Goat secondary antibody (Cat. No. BA-1300)
(which is included in our VECTASTAIN® Universal Kits) is
a cocktail of biotinylated anti-mouse IgG, anti-rabbit IgG, and
anti-goat IgG that recognizes mouse, rabbit, and goat primary
antibodies. This secondary antibody should not be used to
stain rat, mouse, or other rodent, rabbit, goat, bovine, or
sheep tissue due to potential reactivity with endogenous IgG.
3) Choose a convenient format:
Finally, several kits are available in prediluted, stabilized formats
for convenience and ease of use. See kits designated R.T.U. or
Consider Species Cross-Reactivity
When choosing the optimal detection system for your
application, it is important to consider not only the species of
the primary antibody but also the species of the tissue under
examination. If the species of the primary antibody and the
species of the tissue are closely related (e.g. rat and mouse),
the biotinylated secondary antibody may cross react with
endogenous IgG in the tissue section leading to background
staining. The following three options minimize background
staining in these instances:
• Biotinylate the primary antibody (ProtOn™ Biotin Labeling
Kit, Cat. No. PLK-1202) and detect with a Standard
VECTASTAIN® Elite® ABC Kit, PK-6100, (only includes the
• Use a biotinylated secondary antibody specifically adsorbed
to remove cross-reacting antibodies of closely-related species
(e.g. biotinylated anti-mouse IgG, rat adsorbed, Cat. No.
• Use the M.O.M.™ Immunodetection System for applications
of mouse primary antibodies on mouse tissue.
After choosing the VECTASTAIN® ABC Kit for your application,
select a substrate that matches the enzyme system of the kit.
Enzyme Substrates for IHC and ISH