Use of Immunohistochemistry and Confocal Microscopy in the Detection of Adrenergic Receptors
| Abstract |
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With the advent of recombinant protein systems to manufacture large volumes of cloned receptors, antibodies against the α-adrenergic
receptors (α-ARs) have become available (1). These antibodies are useful tools for the immunocytochemical detection of cells containing the adrenergic receptors and
can be used with light-level microscopy. In order to determine whether the receptor protein thus detected is on the cell surface
or is actually contained inside the cell, the technique of confocal microscopy can be applied (for example, see
ref.
2). This powerful technique finds a good application in this case, because double-labeling at the light level can also be done
with relative ease (e.g, see
3,4) for the location of adrenergic receptors in catecholaminergic or serotonergic neurons) compared to asking the same question
at the electron microscopic level. The use of fluorophore-tagged markers for the detection of bound primary antibodies is
central to this method, and with the commercially available variety of these reagents, this becomes a relatively simple procedure.
The procedure consists of a standard immunohistochemical protocol using the primary antibody against the adrenergic receptor
followed by either fluorophore-tagged secondary antibody or indirect-tagged secondary antibody (i.e., biotin-tagged secondary
antibody subsequently visualized with an avidintagged fluorophore). The bound complex including the fluorophore can then be
visualized on laser excitation of the fluorophore at the appropriate wavelength and the subsequent emission of energy at a
wavelength that can be detected by a confocal microscope.
Book Title: Adrenergic Receptor Protocols
Series: Methods in Molecular Biology | Volume: 126 | Pub. Date: Nov-05-1999 | Page Range: 469-473 | DOI: 10.1385/1-59259-684-3:469
Subject: Neuroscience
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