By M. A. Williams, Audrey. M. Glauert
Autoradiography and Immunocytochemistry units out intimately the practise of autoradiographs and strategies of getting ready and use of immunological staining reagents.
This e-book starts off with an advent to the call for for autoradiography and immunocytochemistry and their most suitable use, by way of a dialogue at the immunocytochemistry on the electron microscopical (EM) point on the subject of the guidance of categorized antibodies.
Other subject matters comprise the overall comments on “staining” with antibodies; employing Fer-Ab conjugates to ultrathin sections; and controls in immunoferritin staining. The multistep antibody staining equipment with unmodified proteins; regulate observations in staining experiments utilizing lectins; and development research in EM immunocytochemistry also are lined. this article concludes with an outline of the guidance of electron microscope autoradiographs, together with the radioisotopes in EM autoradiography, coaching of radioactively-labeled tissues and cells, and techniques of utilizing nuclear emulsions.
This ebook serves as an in depth laboratory consultant for researchers and employees to effectively behavior electron microscope innovations.
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Additional info for Autoradiography and Immunocytochemistry: Practical Methods in Electron Microscopy: Autoradiography and Immunocytochemistry Vol 6
The choice of method will depend on the quantities of antibody preparation and other sera available as well as the nature of the equipment the experimenter has. Two methods are outlined here. (i) Examination by immuno-electrophoresis. If the crude conjugate is placed in a well in an agar plate and subjected to electrophoresis, the separated components of the mixture can be allowed to diffuse against serum placed in each of two troughs. These sera would be against peroxidase and the particular IgG or antibody fragments used in the conjugation.
L. (1956), Purification and properties of tobacco ring-spot virus, Phytopathology 46, 60. J. J. Cuculis and J. P. Petrali (1965), Indirect immunouranium technique for staining of embedded antigen in electron miroscopy, Exp. Mol. Pathol. 4, 112. Sternberger, L. , E. J. Donati, J. S. Hanker and A. M. Seligman (1966), Immuno-Diazothio ether-osmium tetroxide (Immuno DTO) technique for staining embedded antigen in electron microscopy, Exp. Mol. Pathol. Suppl. 3, 36. J. Donati and C. E. Wilson (1963), Electron microscopic study on specific protection of isolated Bordetella bronchiseptica antibody during exhaustive labelling with uranium, J.
Nakane and Pierce (1967) describe in detail the preparation of antibodies labelled with horseradis h peroxidase or with wheat-germ acid phosphatase . The acid phosphatase-labelle d antibody has proved too unstable for general use. However, peroxidase is an extremely stable protein and its conjugates are correspondingl y easier to work with. The cytochemica l reaction for peroxidase (Graham and Karnovsky 1966) has the advantage of being widely used. 1. 1 describes methods for preparing peroxidase-labelle d antibodies.