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Optimal conditions for the preparation of ferritin-labeled antibodies defined by binding to their antigen in an ELISA.

Abstract：

:An ELISA with covalently fixed antigens was used to measure and define ferritin-labeled antibodies. In the presented model, RIG covalently fixed to glass tubes served as antigen. Twenty ferritin-labeled SWaRIG conjugates were prepared with different molar ratios of antibody: ferritin: glutaraldehyde. At various dilutions, their ability to react with the antigen was compared. The amount of FRT in the reactive conjugates was measured using alkaline phosphatase-labeled antibodies against ferritin. The covalent binding of antigens to glass surfaces resulted in a low unspecific binding as shown previously. Besides, these glass tubes with antigens immobilized on their inner surface could be used more than once. This aspect of it renders this test particularly suitable for systems where rare or expensive antigens are used. The range of O.D. values reflecting the amount of reactive SWaRIG/FRT detected in 20 different conjugates (dil 1:1000) was spread over a good range which allowed a specification of optimal conditions for FRT labeling of antibodies. It is concluded that in addition to an electron microscopic evaluation of the conjugates, this assay may be very helpful in defining optimal conditions for the coupling procedure.

journal_name

Immunobiology

journal_title

Immunobiology

authors

Rauterberg EW,Schieck C,Kreft H,Römer W

doi

10.1016/S0171-2985(84)80021-2

subject

Has Abstract

pub_date

1984-05-01 00:00:00

pages

439-45

issue

4-5

eissn

0171-2985

issn

1878-3279

pii

S0171-2985(84)80021-2

journal_volume

166

pub_type

杂志文章

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