Primary Antibodies as Key Research Tools
Primary antibodies are key oncology research tools for detecting, measuring, and/or purifying a specific protein or biomolecule. They are immunoglobulins, developed in a range of species (e.g. mouse, rat, rabbit) which bind to your research protein of choice.
Primary antibodies are produced as either monoclonal or polyclonal antibodies, each with their own pros and cons. Primaries have been developed for a wide range of uses, including in vivo studies or basic research applications, or diagnostics.
Monoclonal antibodies recognize an individual epitope per antigen, having been developed from a single B cell parent clone. This makes monoclonal antibodies highly homogenous and therefore specific, providing the perfect antibody for identical production and standardization experiments. Monoclonals also result in no/minimal background staining due to little cross-reactivity.
Disadvantages of monoclonal antibodies include a lack of species cross reactivity, vulnerability to epitope loss after chemical treatment, and sensitivity to changes in experimental conditions.
Polyclonal antibodies, in comparison, are a heterogeneous mix, having been derived from multiple B cell clones. Polyclonal antibodies therefore target several different epitopes of each antigen. As the polyclonal can bind multiple epitopes they can amplify signals from targets with low expression levels and can identify homologous proteins to your target.
Issues with polyclonal antibodies can include variability between batches, cross reactivity, and increased background signalling.
Primary Antibody Applications
In vivo grade antibodies are needed for studies within preclinical models.
For research grade antibodies, there are a variety of applications, with suitability for each use advised for individual antibodies. Uses include:
- Western blot
- IP, etc.
Choosing a Primary Antibody
When you look to choose a primary antibody, you need to take into account the advantages and disadvantages discussed above, as well as what you’ll use it for.
Research Use Only Primary Antibodies
Antibodies for standard research use are available from many vendors. Cell Signaling Technologies and Abcam, to name just two, provide a wide variety of high quality primaries. The list also includes Leinco, RevMAb, ThermoFisher and more.
In Vivo Grade Primary Antibody
For in vivo research, primary antibodies need to be higher purity monoclonal antibodies, often needed in bulk quantities. They should be supplied without preservatives, carriers, or stabilizers in PBS, and have low endotoxin levels.