Localisation of tissue antigens with the fluorescent antibody technique: studies on connective tissue, the kidney and anterior pituitary hormones

Abstract

Problems involved in the use of the fluorescent antibody technique for the localisation of tissue components are discussed.

The need for pure tissue extracts for the immunisation of animals is emphasised and some of the dangers of extraction procedures are described.

The dangers of inadequate preliminary analysis of immune sera are indicated. The need for accurate identification of all antibodies in an immune serum and methods for achieving this are discussed. Appropriate absorptions can be used to get rid of unwanted antibodies and examples axe given of results obtained with such purified antisera.

Measures to counteract the fall in antibody titre caused by conjugation with fluorescein and the presence of adsorbed, rather than chemically conjugated, fluorescein are discussed, including the indirect fluorescent antibody method.

Precautions necessary to establish the specificity of staining with the direct and indirect methods are described. Problems peculiar to studies on auto-antibodies are discussed.

Suggestions are made for further work arising from the present experiments.