Should I use Monoclonal Antibodies or Polyclonal Antibodies?
Both Polyclonal and Monoclonal antibodies each have their own advantages which make them useful for different applications.
Large quantities of polyclonal antibody are relatively quick and inexpensive to produce compared to monoclonal antibodies. They are non specific in that they are capable of recognising multiple epitopes on any one antigen. This capability provides many advantages:
- Can help to increase the signal produced by the target protein as the antibody will bind to more than one epitope
- Less sensitive to antigen changes than monoclonal antibodies
- Useful when the nature of the antigen is unknown
- More robust detection due to multiple epitopes
- Hybridomas provide an immortal cell line with the ability to produce unlimited quantities of highly specific antibodies
- All batches will be identical and specific to just one epitope a particular advantage when manufacturing procedures must be standardised e.g. clinical tests and therapeutic treatments
- The high specificity of monoclonal antibodies decreases background noise and cross-reactivity, helps provide reproducible results and ensures efficiency in affinity purification
Secondary Antibody Overview
Secondary antibodies bind to the primary antibody to assist in detection, sorting and purification of target antigens. To enable detection, the secondary antibody must have specificity for the antibody species and isotype of the primary antibody being used and generally is conjugated. Primary antibodies are typically produced in mouse, rabbit, goat or chicken host species; therefore, anti-mouse IgG, anti-rabbit IgG, anti-goat IgG or anti-chicken polyclonal antibodies are used. Identifying the optimal secondary antibody is normally done based on the nature of the primary antibody and the detection assay. Secondary antibodies are used throughout various types of assays, including ELISA or Western Blot, the secondary antibody type is selected according to the source of the primary antibody, the class of the primary antibody (e.g., IgG or IgM), and the