Conjugation to immunoglobulin

Thefluorochromes and dyes presented here are unconjugated. They maybe conjugated to immunoglobulins (e.g., anti-CD3 ... 

Non-vaccine generated anti-endotoxin reagents that bind and/or neutralize LPS, such as bactericidal permeability increasing protein (BPI), endotoxin neutralizing protein (ENP), and cationic peptide 18 conjugated to immunoglobulin G (CAP 18-IgG) have been developed for the adjunctive therapy of sepsis, but only BPI has progressed to clinical trials (Levin et al., 2000). 

Enzyme Conjugation to Immunoglobulins 4.1. Direct Label Technique... 

O Shannessy, D.J., Doberson, M.J., and Quarles, R.H. (1984) A novel procedure for labeling immunoglobulins by conjugation to oligosaccharide moieties. Immunol. Lett. 8, 273-277. 

Weston, P.D., Devries, J.A., and Wrigglesworth, R. (1980) Conjugation of enzymes to immunoglobulins using dimaleimides. Biochim. Biophys. Acta 612, 40-49. 

A very interesting approach was presented recently by Niemeyer et al. [113]. They prepared covalent DNA-streptavidin conjugates to which biotinylated alkaline phosphatase, beta-galactosidase, and horseradish peroxidase, as well as biotinylated anti-mouse and anti-rabbit immunoglobulins, were coupled. Immobilization of DNA-streptavidin conjugates was performed by hybridization with the complementary oligonucleotides, bound to the surface. It was demonstrated... 

These chemical functions are primarily responsible for metabolic activity besides, all active functions of a small hapten should remain accessible in the hapten carrier conjugate to obtain the most exquisitely specific antibody immunoglobulin (IgG) population of which the immune system is capable,... 

For the procedure, use the conjugated anti-immunoglobulin reagent in Subheading 3., step 11 and go directly from step 13 to step 16. Again, monitoring the DAB development can be critical to a successful result. 

Selected entries from Methods in Enzymology [vol, page(s)j Detection and quantification, 74, 608-616 effect of antigen/anti-body ratio, 74, 613, 614 applications, 74, 614-615 controls, 74, 612 data analysis, 74, 612-613 immunoglobulin G [conjugation to glucose oxidase, 74, 610 heat aggregation, 74, 609-610 heat denaturation, 74, 610 immobilization, 74, 610] precision, 74,... 

The enzymes commonly used as labels in ELISA and other immunochemical reactions include horse radish peroxidase (HRP) and alkaline phosphatase (AP). The enzyme can be covalently coupled to the antibody using glutaraldehyde conjugation to reactive amino groups on the enzyme (lysines) in a phosphate buffered aqueous solution at neutral pH, as shown in Fig. 19 (103). Alternatively, carbohydrates present in the immunoglobulin structure can be cleaved by periodate treatment (see Fig. 20) and bound to free amino groups on the enzyme through a Schiff base reaction (103). 

The CARD approach uses horseradish peroxidase to catalyze the deposition of tyramide molecules that are conjugated to an antihapten immunoglobulin or Streptavidin, which in turn, during the first incubation step, binds to the digoxigenin- or biotin-labeled nucleic acid target. Thus, the number of target sites for the next reaction step is markedly increased. 

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