Product Information
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Aurion EM Reagents
The reagents are prepared using the highest quality antibodies or binding agents available. Antibodies are affinity purified and cross-adsorbed to reduce non-specific reactions.
| Applications | For single and multiple labeling in EM. |
| Size Range | Ultra Small, 6, 10, 15, 25nm |
| Homogeneity : | Coefficient of Variance <12% for 6 and 25nm <10% for 10 and 15nm |
| Multiple Labeling | Choices of size with negligible overlap. |
| Presentation | In PBS (7.6pH) with BSA (1%), and 15mM NaN3 OD520nm = 1 for minimized aggregation |
| Shelf Life | 18 months from production date |
| Specifications | Average size, coefficient of variance, and expiration date is all indicated on the quality control sheet. The activity of each lot is determined using a dot-spot test system. |
| Labeling Capabilities | Large Package Size: 1000-2000 grids Small Package Size: 400-800 grids. |
| Storage | 4-8ºC Do Not Freeze! |
| Particle diameter |
+/ #Au atoms |
+/ MWt. (daltons) |
+/Surface (nm2) |
+/Volume (nm3) |
+/#Particles /ml |
+/#Ab (/part) |
| 6 | 6500 | 1.3.106 | 113 | 113 | 2.4.1013 | 1-2 |
| 10 | 30.103 | 6.106 | 315 | 525 | 5.1012 | 7-12 |
| 15 | 100.103 | 20.106 | 710 | 1770 | 1.5.1012 | 25-40 |
| 25 | 470.103 | 92.106 | 1970 | 8200 | 3.3.1011 | 115-180 |
Ultra Small Reagents
The reagents are prepared using the highest quality antibodies or binding agents available. Antibodies are affinity purified and cross-adsorbed to reduce non-specific reactions.
| Application | EM and LM |
| Size | 0.8nm |
| Presentation | In PBS (7.6pH) with BSA (1%), and 15mM NaN3 |
| Shelf Life | 18 months from production date. |
| Specifications | The activity of each lot is determined using a dot-spot test system. |
| Labeling Capabilities | Large Package Size: 600 slides; 1200 grids. Small Package Size: 240 slides; 400 grids. |
| Storage | 4-8ºC Do Not Freeze! |
| Supplied | Reagents are supplied with 10ml of cold water fish skin gelatin (40%) |
| Particle diameter |
+/ #Au atoms |
+/ MWt. (daltons) |
+/Surface (nm2) |
+/Volume (nm3) |
+/#Particles /ml |
+/#Ab (/part) |
| 0.8 | 15 | 3.103 | 2 | 0.3 | 5.1015 | 0.1-1 |
| 6 | 6500 | 1.3.106 | 113 | 113 | 2.4.1013 | 1-2 |
| 10 | 30.103 | 6.106 | 315 | 525 | 5.1012 | 7-12 |
Why Ultra Small Gold Conjugates?
- More gold particles/secondary antibody instead of more antibodies/gold particle.
- Reduced Steric hindrance.
- More gold particles/surface area in the specimen.
- Maximum penetration.
Reduced Molecular Weight...
- Increased diffusion rates.
- Ultra Small probes behave similar as uncoupled antibodies.
Reduced Probe Size...
- Reduced overall probe size
- Reduced hydrodynamic radius
- Significantly increases penetration into sections and tissues
The Choice for Ligand Receptor Interactions...
- Each Ligand is separate.
- No capping or patching, each probe unit consists of a single ligand.
Universal Probes...
- Optimally suited for high resolution EM, immuno EM, immunohisto at the LM level, and suited for bio-assays as well.
- One probe for all purposes
Aurion R-Gent Silver Enhancement Kits
A. For Light Microscopy and Bioassays
- Light insensitive silver enhancement reagents for increased detection;
the black silver signal does not fade.
-compatible with most counterstains used in light microscopy
-easily detected in bio assays - Easy to use, extremely delayed auto-nucleation
- Used under standard laboratory light conditions.
- Available in: 2x30ml or 2x250ml and consists of a separate Developer and Enhancer.
B. Aurion R-Gent SE-EM- For Electron Microscopy.
These reagents are intended for the enhancement of gold particles, especially Aurion Ultra-Small particles, in electron microscopical visualization of immunodetection and in-situ hybridization experiments. It is our belief that the Aurion R-Gent SE-EM is a significant step forward in the improvement of sensitive detection systems based on Ultra-Small Gold particles.
Features:
- Light insensitive
- Low viscosity
- Time controlled growth of particles
- Easy to use
- High reproducibility
- Low autonucleation
- Available in 30ml size which includes a ready to use enhancer solution, initiator and activator.
Spectrograph displaying the signal increase generated by
silver deposition on Ultra Small gold particles (solid line) vs.
autonucleation (hatched line) with time. Y-axis: optical density
at 415 nm, X-axis: time of enhancement (0-30 min).
Temperature 20ºC.
Recommended on-grid enhancement time is between 15 and 30 minutes
depending on desired particle size.
Aurion R-Gent SE-EM Application Examples
ER exit site in 60 nm-thin cryosection of Hepg2 cells, labeled for
COPII (sec23), detected with F(ab') goat-anti-rabbit, conjugated
to Ultra-Small gold and silver enhanced with R-Gent SE-EM for 30
minutes.
The arrows point to labeled COPII-coats on vesicular and tubular membranes, which are located close to the ER.
The information of a thin section is not sufficient to conclude how the membranes are related to each other- if they are still connected to the ER, or if they are free. Therefore we performed 3D electron tomography on 400nm thick cryosections, which were labeled similar for COPII. (See next picture).
ER=endoplasmic reticulum, PM=plasma membrane, MVB=multi-vesicular body, Bar=100 mikrometer
Courtesy of: Dagmar Zeuschner, Judith Klumperman (Department of Cell Biology, UMC Utrecht,The Netherlands) and Willie Geerts, Abraham Koster (Molecular Cell Biology, Utrecht University,The Netherlands)
 |
2 views of a model of a COPII-labeled ER-exit site, resolved from 400nm thick cryo-sections of Hepg2 cells, labeled like described for the ultrathin section before. Note that the labeling for COPII is assessable throughout the section. ER=light blue. Free membrane carriers of vesicular and tubular shape, partially labeled for COPII=yellow. COPII=silver enhanced-red. Courtesy of: Dagmar Zeuschner, Judith Klumperman (Department of Cell Biology, UMC Utrecht,The Netherlands) and Willie Geerts, Abraham Koster (Molecular Cell Biology, Utrecht University,The Netherlands) |
 |
Membrane labeling with protein A 15 nm gold conjugate. Courtesy of T.Wakefield, University of Auburn, Alabama |
 |
A cross section of dendrite (D) from monkey thalamus, receiving synaptic contacts from three
axon terminals. Terminal t1 is filled with anterograde HRP tracer, and it makes an asymmetric
contact (arrows) onto the dendrite. Terminal t2 is GABA immuno-positive, and it makes a symmetric
contact (arrow heads) onto the dendrite. Rabbit anti-GABA primary antibodies were
labeled with Aurion Ultra-Small gold conjugated goat anti-rabbit secondary antibody and, in
turn, silver enhanced with Aurion SE-EM for 12 minutes.
Courtesy of Mrs. Hong Yi, Dept. of Neurology, Emory University, Atlanta, USA |
 |
A GABA immuno-positive axon terminal from monkey thalamus. Rabbit anti-GABA primary antibodies
were labeled with Aurion Ultra-Small gold conjugated goat anti-rabbit secondary antibody
and, in turn, silver enhanced with Aurion SE-EM for 14 minutes.
Courtesy of Mrs. Hong Yi, Dept. of Neurology, Emory University, Atlanta, USA |
 |
P re-embedding immunogold labeling of parvalbumin in the cortex of mouse brain.
Immunoreactive dendrite (D) with silver enhanced gold particles. Note the uniform shape of particles
with a diameter of about 25nm, about the same as the diameter of microtubules (arrows).
The secondary probe used was Aurion Ultra-Small gold conjugated single Fab goat anti-mouse
(1:100). Enhanced with Aurion R-GENT SE-EM before embedding.
Courtesy of Mrs. Hong Yi, Dept. of Neurology, Emory University, Atlanta, USA |
Desmin labeling using Ultra-Small gold conjugate and silver enhancement. Upper panel:Aurion R-GENT SE-EM, 20 minutes Lower panel: Danscher method, 10 minutes Courtesy of Prof. Dr. J. Müller-Höcker, Pathology, University of Munich, FRG. |
 |
Aurion Gold Tracers
A. ANIONIC GOLD TRACERS
TRACERS for the detection of positively charged compounds. Gold particles (6, 10, 15, 25nm and Ultra-Small) coated with acetylated albumin.
B.CATIONIC GOLD TRACERS
For the detection of negatively charged compounds. Gold particles (6, 10, 15, 25nm and Ultra-Small) coated with methylated albumin.
C. INDIFFERENT GOLD TRACERS
Gold particles (6, 10, 15, 25nm and Ultra-Small) coated with albumin.
- Full package size: 10ml at an OD520 nm of approximately 2.0.
- Reduced package size: 5ml at an OD520 nm of approximately 2.0.
Col-Aurion
For the sensitive staining of proteins in bio assays like immunoblotting.
A ready to use reagent in units of 500 ml, including one vial of 10 ml Tween-20.
VI.
Background Suppression Agents
Background or non-specific reactions may be caused (i) by the proteins adsorbed to the gold particle surface or (ii) by the gold particle characteristics themselves.
A. NORMAL SERUM
Normal rabbit serum is used with the rabbit antibody gold probes, normal goat serum with the goat antibody gold probes, normal sheep serum is used with sheep antibody gold probes and normal monkey serum is used with donkey antibody gold probes etc. to prevent non-specific adsorption of gold probes. Normal Sera are supplied in units of 5ml with 0.01% thiomersal.
B. (I) BOVINE SERUM ALBUMIN (II) COLD WATER FISH SKIN GELATIN
These are used as additives to the incubation buffer which diminish and frequently completely prevent background reactions characteristic to gold particle based reagents. BSA is supplied as a powder in units of 25 grams. Cold Water Fish Skin Gelatin is supplied at a concentration of 40% in units of 10 ml.
C. AURION BSA-C™
This acetylated and linearized form of Bovine Serum Albumin was developed and introduced by AURION in order to deal efficiently with persisting background. By an increased net negative charge and exposure of hydrophobic domains AURION BSA-c™ has a two-fold effect on background. It competes with negatively charged gold particles preventing charge determined background interactions and blocks hydrophobic binding sites (e.g. many plastics used in electron microscopy, hydrophobic plastics used in disposables).AURION BSA-c™ is supplied as a solution of 10% in 30 ml and 100 ml units.
A special AURION newsletter describes in detail the features of AURION BSA-c™. A free copy is available upon request.
D. TWEEN-20
Is a non-ionic surfactant with hydrophobic alkyl side chains. Used at or above the critical micel concentration hydrophobic background is largely prevented. It is supplied in 10 ml units. It is a registered trademark of Atlas Chem. Ind., Inc.
Aurion Blocking Solutions
For EM and LM as well as Bio-assays. Ready-to-use. There is a blocking solution for our entire line of gold conjugates.
Advantages:
- Assurance of the highest consistent quality of the basic components in the solution. All ruminant proteins are tested for the absence of BSE.
- Ready to use and tuned to the specific gold conjugate.
- The solution can be used in other test systems utilizing non-gold particle detection systems.
- No risk of contamination during work.
- Comes in an applicator dropping bottle.
- Solutions available for goat, rabbit, sheep, donkey conjugates as well as for protein A & G in a basic blocking form.
Aurion Gold Sols
Average particle diameters measuring 0.8, 6, 10, 15, 25nm. Unique production procedures for a narrow size distribution and optimum binding properties. Monodisperse, with a coefficient of variance <15% but in most cases <10%.
- SOLS are supplied at an OD520 nm of 1 in 100ml units with detailed instructions for use.
Aurion
ST-US
Ultra-Small atomic gold clusters stabilized with low molecular weight BSA-fragments for covalent binding to (macro) molecules by e.g. glutaraldehyde cross-linking.
- The reagent is supplied in 20ml units with detailed instructions.
