A simple reliable method for producing electron dense markers of uniform size for use in immunoelectron microscopy

A method is presented for the preparation of electron dense markers of defined size for use in immunoelectron microscopy, The electron dense markers were formed by the covalent binding of rabbit IgG and horse spleen ferritin mediated by glutaraldehyde. We demonstrated that the conjugate size, i.e. t...

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Bibliographic Details
Published inJournal of immunological methods Vol. 49; no. 3; p. 237
Main Authors Morris, R E, Saelinger, C B
Format Journal Article
LanguageEnglish
Published Netherlands 26.03.1982
Subjects
Animals
Binding Sites
Chromatography, Agarose
Exotoxins - metabolism
Ferritins - metabolism
Glutaral - metabolism
Horses
Immunoglobulin G - metabolism
Immunologic Techniques
Mice
Microscopy, Electron
Molecular Weight
Pseudomonas - immunology
Rabbits
Rats
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Summary:A method is presented for the preparation of electron dense markers of defined size for use in immunoelectron microscopy, The electron dense markers were formed by the covalent binding of rabbit IgG and horse spleen ferritin mediated by glutaraldehyde. We demonstrated that the conjugate size, i.e. the number of ferritin grains per unit conjugate, was directly related to the time of incubation at 4 degrees C in a 1000 molar excess of glutaraldehyde. Conjugates were separated on the basis of molecular size by Sepharose 4B column chromatography. Three different sized conjugates were used to delineate pseudomonas toxin receptors on the surface of mouse LM fibroblasts; similar numbers of receptors were observed with all conjugates.
ISSN:0022-1759
DOI:10.1016/0022-1759(82)90124-7