Immunocytochemistry with monoclonal anti-immunoglobulin as a developing reagent: application to immunoperoxidase staining and radioimmunocytochemistry.

MacMillan, F; Sofroniew, Michael V.; Sidebottom, E; Cuello, A. Claudio

doi:10.1177/32.1.6418791

Cited by 7 publications

(5 citation statements)

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“…1). Most recently we have developed a radiolabelled monoclonal anti-rabbit IgG (MacMillan, Sofroniew, Sidebottom & Cuello, 1983) and this reagent can be used in an indirect-labelled staining procedure. It thus can provide a radioactive signal for the detection ofconventionally raised rabbit antisera if required (Fig.…”

Section: Monoclonal Antibodiesmentioning

confidence: 99%

Immunocytochemistry and Neurobiology

Cuello¹,

Priestley²,

Sofroniew³

1983

Exp Physiol

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Immunocytochemistry enables the localization of transmitter‐related antigens in tissue sections at either the light microscopic or the electron microscopic level. In the case of neuropeptides and certain transmitters (e.g. serotonin) it has been possible to produce antibodies directed against the putative transmitter itself. In other cases it has not been possible to produce useful antibodies against transmitters but antibodies have been raised against enzymes involved in transmitter metabolism (e.g. tyrosine hydroxylase, glutamic acid decarboxylase) and these antibodies are suitable markers for transmitter systems. Successful immunostaining with an antibody depends on a number of factors, two of the most important being the fixation of the antigen in the tissue and the visualization of the primary antibody once it has bound to the antigen. Techniques available for the visualization of bound primary antibody include the indirect‐labelled immunofluorescence procedure and the unlabelled peroxidase‐antiperoxidase (PAP) procedure. Direct‐labelled immunocytochemistry is not now widely used but is likely to become increasingly important with the introduction of monoclonal antibodies and the development of techniques for the simultaneous localization of multiple antigens. Monoclonal antibody procedures also allow the production of antibodies against antigens which are difficult to purify such as certain transmitter markers (e.g. choline acetyltransferase) and constituents of neuronal membranes. Immunocytochemistry allows the production of detailed maps of the distribution of putative transmitters in the nervous system and in combination with tract tracing procedures is being used increasingly to identify transmitters in neuronal circuits. It has also been important in establishing the transmitter status of various neuroactive compounds in single neurones. Immunocytochemistry can be carried out on post‐mortem samples and is providing information on transmitter distribution in normal and abnormal human brain.

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Section: Monoclonal Antibodiesmentioning

confidence: 99%

Immunocytochemistry and Neurobiology

Cuello¹,

Priestley²,

Sofroniew³

1983

Exp Physiol

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“…One of the immunoreactive boutons is half-embedded in the f'H)DA-labeled peri Stumpf, 1981;Rhodes et al, 1981; or electron microscopic levels (Mohler et al, 1981). The radioautographic detection of labeled primary antibodies or immunoglobulins (Cuello et al, 1982;Hunt and Mantyh, 1984;McLean et al, 1983;Ross et al, 1983;MacMillan et al, 1984; may also be combined with the PAP 35 karyon. Whether both are synaptically linked cannot be ascertained in this plane of section (from Tappaz et al, 1985).…”

Section: Discussionmentioning

confidence: 99%

Characterization of chemically defined neurons and their cellular relationships by combined immunocytochemistry and radioautographic localization of transmitter uptake sites

Bosler

Beaudet

Pickel

1986

J. Elec. Microsc. Tech.

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Radioautography and immunocytochemistry may be combined at the light and electron microscopic levels for simultaneously localizing uptake sites for exogenous transmitter molecules [such as (3H)monoamines or (3H)amino acids] and endogenous transmitter-related antigens (classical transmitters and their synthesizing enzymes as well as neuropeptides) in the central nervous system. Silver grain accumulations indicative of transmitter uptake sites are readily distinguishable from immunocytochemical labels of the peroxidase-antiperoxidase (PAP), avitin-biotin, or colloidal gold methods. The combination of uptake radioautography and immunocytochemistry may be applied to the investigation of (1) the chemical identity of (3H) transmitter-accumulating elements, (2) the coexistence of different neurotransmitters within the same neurons, and (3) the cellular basis of interactions between certain neurotransmitters, in particular monoamines, GABA, and neuropeptides. This article describes and evaluates the method and reviews the available experimental data derived from its application.

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“…This unlabelled staining procedure, described by Sternberger and colleagues, is referred to as the peroxidase antiperoxidase (PAP) procedure. Its sensitivity and specificity has been greatly improved with the development of monoclonal anti-immunoglobulin (MacMillan et al 1984) and antiperoxidase (Cuello et al 1984b, Mason et al 1982, Semenenko et al 1985 antibodies.…”

Section: Apmentioning

confidence: 99%