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Labelled antibodies
Applications such as western blot, immunochemistry or flow cytometry require labelled antibodies
Immunological applications such as ELISA, western blot or immunohistochemistry require the use of conjugates which are covalently linked to the antibody. Some of these conjugates are molecular labels with unique properties enabling their detection through the emission of a measurable signal. Linked to an antibody, these conjugates are useful tools allowing the detection of the protein of interest in your biological sample. Labelling strategies include covalent linking of various bioactive molecules such as biotin, reporter enzymes or fluorophores to the target antibody.
Various enzymes, such as horseradish peroxidase (HRP) and alkaline phosphatase (ALP), can be attached to antibodies and proteins to act as signal-generating molecules via the specific reaction they catalyse.
Other non-enzymatic reporters such as fluorescent labels can be coupled to biomolecules as a means of light generation under excitation at specific wavelengths. The resulting light emission is then easily detected with suitable microscopy equipment, thus allowing the use of multiple labelled antibodies within the same experiment.
Haptens
Coupling also allows small non-immunogenic particles (such as peptides, nucleotides and synthetic or natural compounds) to be carried by larger proteins in order to increase their immunogenicity. Conjugated molecules can thus be used as immunogens when injected into host animals. Such carrier proteins are:
BSA
Bovine Serum Albumin
KLH
Keyhole Limpet Hemocyanin
OVA
Ovalbumin
Correct conjugation methods are essential to direct the specific molecular orientation of the hapten group onto the carrier protein. This orientation is critical for proper presentation and processing in the host animal to ensure the generation of hapten-specific antibodies.