Optimized
Labelling
Optimized Immuno Labelling using AURION BSA-c™ and AURION Blocking Solutions
Newsletter 1 (revised and updated)™
Jan L.M. Leunissen and Peter F.E.M. van de Plas, AURION, Costerweg 5, 6702 AA Wageningen, The Netherlands
Introduction
In any immuno labelling reaction there are at least two components that interact:
Antibodies and markers on the one hand, and specimens with small amounts of antigen on the other.
A number of different interactions may occur between antibodies and specimens, leading to the final results:
(i) specific reactions
They result from the interaction between the antibody cascade and the antigens. Successful labelling depends on the quality of the specific binding agent and the preservation and availability of antigen(s). Fixation, denaturation and masking of antigens play a major role. Antibodies may also recognize areas closely resembling epitopes in molecules other than the antigen under investigation. As undesirable as such reactions may be they are nevertheless based on specific interactions. Therefore, strictly speaking, considering such reactions as background is not correct.
(ii) background reactions
Strictly, background is the result of unintended “non-specific” reactions. They are governed by general physical chemical properties of both the specimen/substratum and of the primary antibodies and secondary antibody/marker conjugates.
Is it possible to have one without the other? Yes, but it is certainly not the rule. Manufacturers of immunoreagents use proprietary methods in an attempt to produce labelled secondary antibody reagents with minimized stickiness. But that is only one side of the story: the researcher is confronted with the characteristics of the specimen as well and will thus be interested in the answer to the question:
“Which incubation conditions will promote specificity and reduce background in the best achievable way in my specimens?”
AURION, being a research-dedicated company, has diligently looked into the principles of specificity and background, unraveling their common denominators and differences. From the vast range of fluorescent markers via enzyme conjugates to particulate labels, the products resulting from these R&D activities provide unparalleled signal-to-noise ratios
REGARDLESS OF THE MARKER SYSTEM
This is achieved by the combined application of AURION Blocking Solutions and the unique incubation solution additive AURION BSA-c™. The Blocking Solutions guarantee an effective and lasting specimen blocking prior to antibody exposure. Incubation with antibodies and labels as well as washing are done using AURION BSA-c™ which acts where it matters: preventing non-specific binding without affecting antibody-antigen interactions, interfering only where antibodies are inclined to stick to specimens.
