Immunoglobulins conjugation

Hurwitz, E., Maron, R., Arnon, R., Wilchek, M., and Sela, M. (1978) Daunomycin immunoglobulin conjugates, uptake and activity in vitro. Eur. J. Cancer 14, 1213. 

Sodium azide inhibits peroxidase enzyme, therefore it should be left out from the diluents used with PO-immunoglobulin conjugates. Add 0.05% Tween-20 when diluting gold reagents. 

Alkaline Phosphatase-Immunoglobulin Conjugate (Glutaraldehyde Protocol)... 

The sections are washed in PBS for 5 min and incubated for 30 min in fluorescein isothiocyanate (FICT)-labeled swine antirabbit immunoglobulin conjugate at 1 20 dilution in PBS. After being washed in PBS for 5 min, the sections are mounted with an aqueous mounting medium. The sections are observed under an epiilluminating fluorescent microscope. The negative control is not incubated with the primary antibody, and a frozen section from a patient with the known condition is used as the positive control. 

Rabbit antimouse immunoglobulin conjugated to horseradish peroxidase (RbaMIgHRP). Store at 4°C. 

Second andbody (Dako) Goat andrabbit immunoglobulins conjugated to horseradish peroxidase. If the primary andbody is the mouse monoclonal andbiodn, the second andbody should be rabbit andmouse immunoglobulins (Dako). 

Ghosh MK, Kildsig DO, Mitra AK. Preparation and characterization of methotrexate-immunoglobulin conjugates. Drag Des Deliv 1989 4 13-25. 

Yokoyama, M., Okano, X, Sakurai, Y., Kataoka, K. and Inoue, S. (1989b) Stabilization of disulfide linkage in drug-polymer-immunoglobulin conjugate by microenvironmental control. Biochem. Biophys. Res. Common., 164(3), 1234-1239. 

ELISA (Engwall and Perlman 1971) uses antibody systems able to release a chromogen substance. The sandwich technique is the most commonly used variant. The Ag is first coated on a solid surface - often, plastic wells of a microplate. The antibodies (Ah) present in the serum are then bound with the Ag. Finally, the Ab-Ag complex is both bound with an anti-human immunoglobulin conjugate able to bind with it and combined with an enzyme, which when it reacts with a chromogenic substrate, releases a coloured compound. The intensity of coloration, which is proportional to Ab concentration, is read by a spectrometer. Results are expressed as an optical density or with a reference to a pooled sera and expressed as arbitrary units (Fig. 3.5.5). 

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