Conventional Immunogold Reagents are
available in four size classes. The monodisperse size population makes the
conjugates suited for multiple labeling with no overlap. The Conventional
Immunogold Reagents are the classical conjugates in immuno electron microscopy;
they are a good choice when the antigen is abundant and the accessibility of the
antigen is relatively good.
Introduction
The conventional labeling approach in transmission
and scanning electron microscopy utilizes secondary immunogold reagents based on
particles that can be observed without enhancement. These conjugates are suited
for single and multiple labeling in electron microscopy, when the number of
antigens available for binding is such that a relevant signal can be obtained.
The AURION Conventional Immunogold Reagents are built around colloidal gold
particles with sizes of 6, 10, 15 or 25 nm. The particle population is
monodisperse and thus shows minimal size variation and overlap. Typically, the
coefficient of variance for the 6 and 25 nm particle size conjugates is less
than 12%, whereas the 10 and 15 nm size conjugates show less then 10%
variation.
The table below lists a few physical characteristics of gold
conjugates.
Particle
#Au MWt.
#particles/ #Ab/
diameter atoms
(daltons)
ml
(part.)
6
nm 6500
1.3·10^6 2.4·10^13
1-2
10 nm 30·10^3
6·10^6
5·10^12 7-12
15 nm
100·10^3 20·10^6
1.5·10^12 25-40
25 nm
470·10^3 92·10^6
3.3·10^11 115-180
Product
Description
AURION Conventional Immunogold Reagents are tailored to
contain 10-20 µg of specific protein/ml. The reagents are supplied in PBS
with 1% Bovine Serum Albumin and 15 mM NaN3 at an OD520nm of 1.0 to warrant
prolonged cluster free storage. The activity of each lot is determined using a
dot-spot test system as described by Moeremans et al., J. Immunol. Methods, 74,
(1984), 353. Actual lot specifications (size, variation and expiry date) are
indicated on the accompanying package insert.
Regular package: for the
labeling of 1000-2000 grids
Small package: for the labeling of
400-800 grids
Application Instructions
On-Grid Labeling
For most applications grids are floated on
top of drops of dilute reagent displayed on a sheet of Parafilm™. The use
of gold or nickel grids is recommended.
Whole mount and intact cell
labeling
Specimens are kept floating in dilute reagent on a rocking
table.
Recommended Incubation Solution:
PBS,
(10 mM Phosphate
buffer, 150 mM NaCl),
0.1-0.2 % AURION BSA-C™
15 mM NaN3
pH
7.4
Specificity
Aurion offers the widest range of Conventional
Immunogold Reagents. Please refer to the list of available reagents under
‘Ordering Information’.
AURION Conventional Immunogold Reagents
are prepared using the highest quality antibodies or binding agents available.
All antibodies are immuno affinity purified and immuno cross-adsorbed to reduce
non-specific reactions.
Information in the reactivity of Protein A and
Protein G can be found in the section "Sharing our knowledge" or by following
this link.
Storage
AURION
Conventional Immunogold Reagents 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.
Immunolabeling of the periplasmic space in
ultrathin cryosections of Escherichia coli with a protein A gold
conjugate.