Obtaining Specific Antibodies

By immunizing animals with specific hapten-carrier conjugates, we have been very successful in obtaining specific antibodies to nicotine, cotinine (17-19), y-(3-pyridyl)-Y-oxo-N-methylbutyramide (18), N -nitrosonomicotine (42), the nicotinamide nucleotide analogues of nicotine and cotinine (43) and, most recently, the intermediate formed during the nicotine to cotinine conversion (Figure 1) (44). We have failed, despite many attempts, to obtain antibodies to another metabolite of nicotine, i.e. nicotine-N -oxide. The hapten derivatives may have been... 

Alkaline phosphatase assays based on 3-glycerophosphate now appears to be obsolete, and methods buffered by either glycine or barbital are also obsolete as these buffers inhibit ALP or are poor buffers. Serum alkaline phosphatase is known to be composed of several isoenzymes which presumably arise from bone, liver, intestine, and placenta. The placental alkaline phosphatase is known to be much more resistant to heat denaturation than the other isoenzymes, and this resistance provides a simple test for it (5). The other enzymes can be separated through the differential inhibition by phenylalanine, by electrophoresis and by specific antibodies. However, the clinical usefulness of the results obtained is in doubt (23). 

Immuno-histochemical staining of intracellular bacteria in filarial nematodes has been obtained using antibodies against GroELand catalase (Henkle-Duhrsen etal., 1998 Hoerauf etal., 1999) the specificity of these antibodies is unknown, but it is expected to be low because both GroEL and catalase show high level of amino acid conservation throughout the proteobacteria. nd = not done. 

The requirement for high-specificity antibodies cannot be understated. Testing the antibodies for specificity is an important aspect of any immu-nocytochemical study. Antibodies used for immunocytochemical assays may be developed especially for the localization project, or more likely developed for other aspects of the project or obtained from other investigators. Sharing probes such as antibodies is now widespread and indeed an expectation for publication in most journals. Antibodies obtained... 

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,... 

With individuals of blood group 0, injection of A and B substances gave a big increase in the amounts of precipitins. For example, the specific antibodies to the blood group A substance could be obtained from the specific precipitate by the use of 15% sodium chloride for dissociating... 

A specific antibody against COX-2 and CA2 were obtained from Santa Cruz Biotechnology, Inc ( sc-19999 and sc-25596), respectively. 

In general, most of published immunochemical methods on chloramphenicol assay are based on antibodies particularly specific for the aromatic ring and the propanediol moiety of chloramphenicol. This is due to the fact that the main chloramphenicol analogues such as thiamphenicol differ from chloramphenicol in one of these parts of the chloramphenicol molecule. To obtain such antibodies, chloramphenicol was linked to the carrier bovine serum albumin at the acyl chain of the molecule by mixed anhydride or carbodiimide reactions (49, 50). In both... 

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