AURION IMMUNOGOLD REAGENTS EDITION VI PRODUCT INFORMATION
NEW!...
The successor to R-Gent SE-LM with improved signal contrast, reaction speed, and reproducibility!
zzz R-Gent SE-Intense Silver Enhancement Reagents
The Immunogold Silver Staining technique (IGSS) finds application both at the electron microscope and the light microscope level. In addition the technique is used to identify proteins or nucleic acids after blotting.
Light microscopical and macroscopical visualization of the latent gold signal requires an enhancement system that renders a high contrast signal. Light insensitivity and negligible autonucleation are required for ease of handling and low background. AURION R-Gent SE-Intense is a two-component reagent that meets these requirements.
Introduction
The silver enhancement reaction is based on the gold parti- cle catalyzed reduction of Ag+ to metallic silver using pho- tographic developing compounds as the electron source. For light microscopy and immunoblotting applications the gener- ated silver signal should be of high contrast. Furthermore the signal should be permanent and compatible with coun- terstaining.
Aurion R-Gent SE-Intense is the successor of Aurion R-Gent SE-LM. Key aims during the development of this new prod- uct were: contrast, reaction speed and reproducibility.
Product Description
Aurion SE-Intense is a two component reagent consisting of a Developer and an Enhancer. When combined they consti- tute a Silver Enhancement Reagent which increases the average gold cluster or particle size by deposition of metallic silver facilitating detection at the light microscopical level. The generated brown-black signal is compatible with most counterstaining methods.
The intensity of the signal facilitates detection of positive signals in bioassays. AURION R-GENT SE-Intense has been tailored for the enhancement of AURION Ultra Small Immunogold reagents and Aurion 2nm gold reagents is however equally suited for silver enhancement of the larger sized particles in the AURION Conventional Immunogold reagents.
For enhancement, equal amounts of the DEVELOPER and ENHANCER are mixed well and applied to the specimen. The enhancement mixture is easy-to-use, exhibits extremely delayed auto-nucleation and can be used under standard laboratory light conditions. Typical enhancement times are between 15 and 25 minutes. Auto-nucleation becomes visi- ble only after 40 minutes. Light microscopical specimens may be counterstained according to standard procedures. The enhancement mixture has a pH value of 8.2-8.6. AURION R-GENT SE-Intense is available as a kit in two unit sizes (2 x 30 ml or 2 x 250 ml) and consists of a separate DEVELOPER and ENHANCER. The supplied amounts acco- modate 600 and 5000 LM specimens respectively at 100 µl/specimen, or 60 and 500 bio assay specimens at 1 ml/specimen. The reactivity is tested on dot-spots and the absence of autonucleation is monitored by spectrophotomet- ric techniques.
Bright field light microscopy: Detection of actin on paraffin sections of formalin fixed rat kidney tissue using AURION GAM IgG Ultra Small and AURION R-Gent SE-Intense silver enhancement reagent
TECHNICAL TIPS...
Application Instructions Detailed information is provided in the package inserts.
Electron Microscopy
Post-embedding enhancement (on-grid) Place grids on drops of the enhancement mixture arrayed on a sheet of Parafilm™.
Pre-embedding enhancement
Incubate specimens (e.g. vibratome sections) in Petri dishes or Eppendorf tubes containing the enhancement mixture, with agitation. When enhancement is completed, the specimens are washed with distilled water. A fixation step with photographic fixer is not required.
Light Microscopy On-slide or -coverslip Labeling
A few drops of the freshly prepared enhancement mixture are applied to cover the specimen. During enhancement the specimens are kept in a moist chamber. The ongoing enhancement may at intervals be monitored with an inverted light microscope. When enhancement is judged to be complete, the specimens are washed with distilled water. A fixation step with photographic fixer is not required.
Whole Mount Labeling
Specimens like floating (vibratome) sections can be enhanced in Petri dishes or 6-24 well plates.
Bio Assays
Light microscopy evaluation of tubulin labeling with Ultra Small Immunogold Reagents and silver enhancement.
Upper panel: bright field mode Lower panel: epi-polarization mode
Depending on the type of assay the enhancement for bio assays can be performed in sealed plastic bags, Petri dishes or in disposable screw cap sealed tubes.
Storage
The AURION R-GENT SE-Intense components are stored at 4°C and allowed to reach room temperature before use.
Additional Information
For additional information, please see the Aurion Immunogold Newsletters and Technical Tips on our website.
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