Primary antibodies are key reagents used to detect or quantify particular antigens across various sample types.

Polyclonal antibodies are a heterogeneous mixture of antibodies that recognize different epitopes of a target antigen. Targeting multiple epitopes on the same antigen can enhance sensitivity, allowing for the detection of low-abundance targets or lower concentrations of the antigen. However, polyclonal antibodies can exhibit batch-to-batch variability, cross-reactivity, and lower specificity, which can be mitigated by cross-adsorption to remove antibodies that target different antigens that may be present.

Monoclonal primary antibodies overcome limitations often encountered with other production methods as they consist of antibodies with identical variable regions. Recombinant monoclonal antibodies build on this further by eliminating genetic drift that may result in variability in the final product. Moreover, large-scale production of antibodies against challenging targets like toxins, nucleotides, and membrane-bound proteins can be achieved when using recombinant monoclonal antibodies.

In cases where polyclonal antibodies are traditionally used, such as for analyzing low-abundance targets or detecting multiple post-translational modifications, recombinant multiclonal antibodies provide an ideal alternative. These antibodies are a defined mixture of carefully selected recombinant monoclonal antibodies, each recognizing different epitopes on the same antigen, delivering exceptional sensitivity, specificity, and reproducibility not achievable with conventional polyclonal antibodies.

We also offer matched antibody pairs - a pair of antibodies confirmed to bind to different epitopes. These are suitable for applications like ELISA, where a difference in the epitope recognized by the different antibodies is crucial for accurate results.

Our chimeric antibodies are engineered antibodies using the same Fc region (the binding/variable region of the antibody) and replacing the backbone region with that of a different species. By doing so, you are able to use the same antibody in different model systems for easier data comparability and you can eliminate issues of species-on-species interactions.

Our fluorescently conjugated primary antibodies deliver bright and reliable results for immunofluorescence and flow cytometry. These fluorescently labeled antibodies simplify protein analysis, saving valuable time and resources by eliminating the need for a secondary antibody. We offer a wide and ever-growing selection of fluorescently conjugated antibodies, ideal for several applications. We also offer fluorescently conjugated antibody panels for multiplex-based applications.

Similarly, we offer a range of antibodies with other conjugates, such as tags or enzymes suitable for several applications, including signal visualization in tissues, pull-down assays, and quantitative measurement of your target in a range of samples.

Additionally, our carrier-free antibodies are optimized for antibody labeling. These antibodies can be added directly to the conjugation reaction without the need for purification, eliminating antibody loss and bypassing the need for buffer exchange or antibody concentration. When paired with our Lightning-Link® labeling technology, conjugation becomes even more straightforward and efficient. Using our unconjugated and carrier-free antibodies, you can develop your own panels of fluorescent antibodies, thus providing you with an additional level of flexibility when designing your experiments.

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FAQs

How are primary antibodies used in research?

Primary antibodies are used in research to specifically bind to and detect target antigens within various biological samples, facilitating the localization, quantification, and analysis of proteins. These antibodies are integral to techniques such as western blotting, immunohistochemistry, ELISA, and flow cytometry, enabling researchers to study protein expression, localization, and interactions. Commonly, a secondary antibody is used to detect the primary antibody and allow for signal amplification, which is particularly valuable when working with low-abundance targets.

What is the difference between primary and secondary antibodies?

Primary antibodies are designed to bind directly to a specific antigen, whereas secondary antibodies bind to primary antibodies, often conjugated to a reporter molecule that enables detection or visualization in various assays. The use of a secondary antibody allows for signal amplification, as multiple secondary antibody molecules can bind to one primary antibody, and each secondary antibody will carry a reporter molecule, meaning multiple reporter molecules per one primary antibody molecule.

Conjugated primary antibodies are antibodies that have been directly labeled with a detectable tag or reporter molecule, such as a fluorescent dye, enzyme, isotope, nanoparticle, or another tag, eliminating the need for a secondary antibody for detection. They simplify experimental workflows by directly binding to the target and providing a detectable signal, saving time and reducing complexity. However, by using both primary and secondary antibodies, you can amplify low-abundance signals, thus increasing the sensitivity.

How do primary antibodies bind to antigens?

Primary antibodies bind to antigens through their antigen-binding sites within the variable regions. These specifically recognize and attach to unique epitopes on the antigen via noncovalent interactions such as hydrogen bonds, electrostatic forces, and hydrophobic interactions.

What is the role of primary antibodies in ELISA?

In indirect and competitive ELISA, primary antibodies specifically bind to the target antigen to capture it, thereby forming an antigen-antibody complex.