AURION IMMUNOGOLD REAGENTS EDITION IV PRODUCT INFORMATION
zzz Conventional ImmunoGold Reagents
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 sec- ondary 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 anti- gens available for binding is such that a relevant signal can be obtained.
TECHNICAL TIPS... Application Instructions
Detailed information is provided on the package inserts.
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
Additional Information
For additional information please see the Aurion Immunogold Newsletter and Protocol sections
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
Diameter 6 nm
10 nm 15 nm 25 nm
Features
z For single and multiple labeling z Gold particle sizes of 6, 10,15 and 25 nm z Monodisperse particle population
z OD520 nm of 1.0 to warrant cluster free stor- age using a dot-spot test system.
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 speci- fications (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
Specificity Aurion offers the widest range of Conventional Immunogold Reagents.
AURION Conventional Immunogold Reagents are prepared using the highest quality antibodies or bind- ing agents available. All antibodies are immuno affinity purified and immuno cross-adsorbed to reduce non-specific reactions.
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. Courtesy M. de Jong
z Minimal size variation and overlap
z Coefficient of variance: <12% for the 6 and 25 nm conjugates and <10% for the 10 and 15nm conjugates
+⁄– #Au atoms
6500/td> 30·103
100·103 470·103
+⁄– MWt. (daltons) 1.3·106 6·106
20·106 92·106
+⁄– #Particles /ml
2.4·1013 5·1012
1.5·1012 3.3·1011
+⁄– #Ab (/part) 1-2
7-12 25-40 115-180
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