[28] Use of cell-specific monoclonal antibodies to isolate renal epithelia (original) (raw)

Methods in Enzymology, 1989

Abstract

Publisher Summary This chapter describes the immunoadsorption procedures for isolating large, homogeneous populations of both canine and rabbit collecting tubule cells. Cultures of these cells exhibit a number of the differentiated properties of the parent cells even after several passages. In principle, immunoadsorption should be applicable to isolating any cell type against which a specific antibody is available. The procedures for isolating canine and rabbit cortical collecting tubule cells—canine cortical collecting tubule cells and rabbit cortical collecting tubule cells—are described. Unless one already has available antibodies specific for a cell surface protein on the cell of interest, it is necessary to prepare monoclonal antibodies. For immunodissection, the nature of the antigen is of secondary importance, but it must be an ectoantigen present in relative abundance and only on the cell of interest. It is also important that the antibody be of the immunoglobulin G class because IgGs are more stable and easier to purify. Indirect immunofluorescence is used to test for antibodies to specific renal cell types.

William Spielman hasn't uploaded this paper.

Let William know you want this paper to be uploaded.

Ask for this paper to be uploaded.