Blocking Solutions
The signal-to-noise ratio determines the
quality of any detection experiment. This ratio is in principle determined by
the characteristics of specimen and detection reagents. With adequate blocking
and incubation media, background reactions are minimized whereas specific
reactions are not hampered.
Aurion has developed Blocking Solutions
which effectively block ‘sticky’ surfaces by multi-point
hydrophobic and charge-based interactions. The Blocking Solutions are
specifically designed to meet the characteristics of each type of secondary
conjugate.
Introduction
The AURION Blocking Solutions are used to prevent
immunoreagents from binding non-specifically to specimens with
“sticky” surface properties.
Procedures to eliminate
background comprise three main steps:
- To suppress residual aldehyde
activity
- To saturate multipoint hydrophobic moieties and positive
charges with high molecular weight compounds such as those present in the
AURION Blocking Solutions
- To reduce aspecific binding of immunoreagents
caused by hydrophilic interaction with competing molecules in the incubation and
washing solution. AURION BSA-c™ is a particularly effective reagent for
this purpose.
These steps should be balanced for optimum
results.
Product DescriptionAURION Blocking Solutions are prepared
using specially selected compounds. All ruminant proteins are obtained from
healthy livestock.
AURION Blocking solutions contain Bovine Serum
Albumin and Cold Water Fish Skin Gelatine in phosphate buffered saline with
sodium azide as preservative. Normal serum may have been added as indicated on
the label. The blocking capacity of each lot is determined using a dot-spot
test system as described by Moeremans et al., J. Immunol. Methods 74, (1984),
353.
Each package contains 30 ml of solution. It accommodates 300 specimens
for light microscopy at 100 µl/specimen (~ 3 drops), or 1000 EM grids at
30 µl/specimen (~ 1 drop).
Application
InstructionsDetailed information is provided on the package
inserts.
Electron MicroscopyOn-Grid Labeling
Placing grids on top of drops of
Blocking Solution arrayed on a sheet of Parafilm™ after they have been
treated for aldehyde inactivation.
Pre-embedding Labeling
Floating specimens in Blocking
Solution, preferably on a rocking table.
Light
MicroscopyOn-slide or
-coverslip Labeling
Applying a few drops of Blocking Solution
to cover the specimen.
Bio
Assays
Incubating specimens (e.g. immunoblot strips) in
Blocking Solution on a rocking table in screw-cap sealed disposable
tubes.
Specificity
Blocking Solutions are available in the following
specificities:
- serum-free: for use with Protein A and Protein G
Gold conjugates.
- with Normal Goat serum: for use with reagents based on
secondary antibodies raised in Goat.
- with Normal Rabbit serum: for use
with reagents based on secondary antibodies raised in Rabbit.
- with
Normal Sheep serum: for use with reagents based on secondary antibodies raised
in Sheep.
- with Normal Donkey serum: for use with reagents based on
secondary antibodies raised in Donkey.
StorageAURION Blocking
Solutions have a guaranteed shelf life of 18 months from the date of quality
control analysis.
The products should be stored at 4-8°C. Freezing is
not recommended.
© Aurion 2007 - 2010