For your convenience Aurion has developed Ready-To-Use Incubation buffer solutions, with long term stability, that significantly reduce the risk of background. The main buffer component is AURION BSA-c™, a unique buffer additive that prevents aspecific binding of antibodies and immunodetection reagents to polycationic sites in the specimen like histone proteins in the nucleus.
The Ready-To-Use Incubation buffer solutions are applicable in all types of immuno(cyto)chemical procedures, not only in ImmunoGold Silver Staining, also in immunofluorescence and immuno-enzymatic techniques in microscopic and macroscopic applications.
AURION Ready-To-Use Incubation buffers: Solutions to success in your immuno(cyto)chemical procedure!
In immuno(cyto)chemistry a good signal to noise ratio is the key to success. To obtain low background levels, non-specific binding to the ‘sticky’ surfaces in the specimen, by multi-point hydrophobic interactions between the specimen and immunoreagents, needs to be circumvented. There are two ways to achieve this. The first option is the use of blocking proteins that physically occupy non-specific binding sites. This blocking step is applied prior to the incubation with primary antibody. Alternatively non-ionic detergent molecules, which prevent immunoreagents to bind to hydrophobic compounds in the specimen, can be used. These detergents need to be present during the entire immunodetection method.
During the incubation with immunoreagents, electrostatic and ionic interaction between the specimen surface and immunoreagents needs to be controlled to eliminate charge determined background. AURION BSA- c™ has the unparalleled ability to effectively prevent this type of non-specific staining. The activity of AURION BSA- c™ is warranted between pH 7.2 and 7.6. A special newsletter on the prevention of background staining in immunochemistry can be found here.
Aurion has developed two types of ready-to-use Incubation buffers, based on AURION BSA-c™, that can be implemented in most immuno(cyto)chemical procedures. These Incubation buffer solutions are used throughout the whole set of reaction and washing steps, i.e., from immediately after the blocking step (when appropriate), through the primary antibody incubations, in-between washing steps, secondary incubation and the final washing steps.
Aurion Ready-to-use Incubation buffer solutions have a guaranteed shelf life of 18 months.
A switch towards a different buffer system after the immuno incubation is optional. For example to a Tris buffer at pH 8-9 for the optimal development of the alkaline phosphatase signal.
AURION Ready-to-use incubation buffer solutions based on AURION BSA-cTM are available in two buffer compositions:
PBS (10mM Phosphate buffer, 150mM NaCl) + 0.1% BSA-c™ + 15mM NaN3, pH7.4
PBS (10mM Phosphate buffer, 150mM NaCl) + 0.1% BSA-c™ + 0.1% TWEEN-20® + 15mM NaN3, pH 7.4
900.099-PBS: The ready-to-use Incubation buffer that can be used in all immuno (cyto)chemical methods. A protein block step is required. The use of 900.099-PBS is recommended for those specimens and applications were (ultra)structure is important and preservation of structure and/or extraction is not warranted by the specimen preparation method (e.g., for Tokuyasu cryo sections).
900.099-PBST: A ready-to-use Incubation buffer that combines the positive effect of both AURION BSA-cTM and Tween 20® with respect to reduction of charge determined and hydrophobic background This incubation buffer solution suits those applications were preservation of structure is less important. A protein block step prior to the actual incubation may not be required.
Detailed information is provided in the package insert
AURION Ready-to-use Incubation buffer solutions 900.099-PBS and 900.099-PBST have a guaranteed shelf life of 18 months from the date of quality control analysis. The buffers should be stored at 4-8°C. Freezing is not recommended.