How Antibodies are Produced
Specific antibodies are produced by injecting an antigen into a mammal, such as a mouse, rat, rabbit, goat, sheep, or horse. Blood isolated from these animals contains polyclonal antibodies — multiple antibodies that bind to the same antigen — in the serum, which can now be called antiserum. Antigens are also injected into chickens for generation of polyclonal antibodies in egg yolk. To obtain an antibody that is specific for a single epitope of an antigen, antibody-sereting lymphocytes are isolated from the animal and immortalized by fusing them with a cancer cell line. The fused cells are called hybridomas, and will continually grow and secrete antibody in culture. Single hybridoma cells are isolated by dilution cloning to generate cell clones that all produce the same antibody, known as monoclonal antibodies. Polyclonal and monoclonal antibodies are often purified using Protein A/G or antigen-affinity chromatography.
Antibodies in Research
Purified antibodies are used in many applications. They are most commonly used to identify and locate intracellular and extracellular proteins. Antibodies are used in flow cytometry to differentiate cell types by the proteins they express. Different types of cell express different combinations of cluster of differentiation molecules on their surface and produce different intracellular and secretable proteins. They are also used in immunoprecipitation to separate proteins and anything bound to them (co-immunoprecipitation) from other molecules in a cell lysate, in Western blot analyses to identify proteins separated by electrophoresis, and in immunohistochemistry or immunofluorescence to examine protein expression in tissue sections or to locate proteins within cells with the assistance of a microscope. Proteins can also be detected and quantified with antibodies, using ELISA and ELISPOT techniques.
Custom Research Antibodies
ARP American Research Products is a custom antibody supplier and one of the premier antibody supplier companies for leading academic and biotechnology firms. In addition to our extensive catalog of Monoclonal and Polyclonal Antibodies, we can create custom antibodies if your research calls for specific requirements. We invite you to contact us to discuss how ARP's custom antibody services can support your ongoing research efforts.
Contact us (800) 832-2611 or email at firstname.lastname@example.org for custom antibody services and for assistance in selecting from our antibody selection.