ELISA using

Uto, I., Ishimatsu, T., Hirayama, H., Ueda, S., Tsuruta, J., and Kambara, T. (1991) Determination of urinary Tamm-Horsfall protein by ELISA using a maleimide method for enzyme-antibody conjugation. J. Immunol. Meth. 138, 87-94. 

Firefly luciferase, together with luciferases from other organisms, can be used as the labeling enzyme in immunoassays and nucleic acid assays [25], Recently, a highly sensitive BL ELISA using firefly luciferase was applied to thyreo-... 

Other attempts have been made to detect BPA at a low concentration range. Thus Kodaira et al. [274] analyzed BPA in urine samples with an assay that showed a working range between 0.5 and 5 pg L The assay was validated by HPLC. DeMeulenaer et al. [275] developed an indirect competitive ELISA using PAbs obtained from chicken egg yolk, but the assay achieved an IC50 value of only 570 pg L-1. 

Comparison of the three techniques (LC-MS, LC-FL and ELISA) used to determine NPEO leads to the conclusion that LC-MS is the most... 

Vandenbroeck et al.7 used an ELISA to determine the recovery of immu-noreactive porcine interferon-gamma (IFN-y) from E. coli inclusion bodies. The ELISA used a polyclonal coating antibody with detection by a MAb. The inclusion bodies were solubilized in diluted 6 M guanidine/HCl and IFN subsequently refolded by its removal. The antiviral activity of the interferon was measured with a bioassay using the cytopathic effect (CPE) of vesicular stomatitis virus (VSV) on bovine kidney cells. The results of this study showed that the immu-noreactivity measured by ELISA matched the biological activity measured by bioassay. 

The ELISA uses equipment that is commonly accessible in immunochemical laboratories. They are usually formatted using microtiter plates, which have 96 wells. Depending on how many wells are used for standards and controls, many test samples can be run on one plate. It is common practice to run multiple plates... 

After incubation, the supernatants of the ceU cultures can be used for the cytokine assay by capture enzyme-linked immunosorbent assay (ELISA) using specific antibody pairs and recombinant cytokine standards and ELISA plates. 

Apart from radioimmunoassays, various enzyme-linked immunosorbent assays have been described as well. Campbell et al. (42) first reported a sensitive and specific ELISA using polystyrene tubes and a polyclonal antibody. However, the performance of this method was not evaluated with real samples but only with standards and aqueous muscle tissue extracts. Sensitive ELISAs were also developed for the determination of chloramphenicol in milk (43) and eggs (44) the results drawn by the latter assay correlated well with those obtained by application of a radioimmunoassay. 

Yuesheng, L., Rabello, A.L.T., Simpson, A.J.G. and Katz, N. (1994) The serological differentiation of acute and chronic Schistosoma japonicum infection by ELISA using keyhole limpet haemocyanin as antigen. Transactions of the Royal Society of Tropical Medicine and Hygiene 88, 249-251. 

Deng, A., M. Franbk, and V. Kolaf (1999). Determination of atrazine in soil samples by ELISA using polyclonal and monoclonal antibodies. Food Agric. Immunol., 11 135-144. 

Roberts, C.J. and L.S. Jackson. 1995. Development of an ELISA using a universal method of enzyme-labelling drug-specific antibodies Part I Detection of dexamethasone in equine urine. J. Immunol. Methods 181 157-166. 

Fig. 1. ELISA using a second antibody with an attached (conjugated) enzyme that converts a colorless substrate to a colored product.

Corporation, Lansing, MI) is a sandwich ELISA used for the quantitative analysis of minimally processed soy flour protein in food products such as cookies, crackers, chocolate bars, and cereals in the range of 2.5-25mg/kg. 

Scheme 4.92 Assay scheme of a micro-ELISA using antibodies immobilized on PS beads. Reprinted with permission from [403], Copyright 2004 The Royal Society of Chemistry.

One week after the third or fourth boost, antisera may be collected and tested for immunoreactivity by ELISA using free peptide (see Subheading 3.3.1.) or by flow cytometry using receptor transfectants, cell lines or leukocytes known to express the receptor (see Subheading 3.3.2.),... 

Indirect ELISA Using Antibody-Coated Plates... 

Chlorination products (mainly chlorotyrosine) are measured by HPLC (Fig. 8) or gas chromatography-mass spectrometry. Also, 3-nitrotyrosine can be detected in protein hydrolysates by HPLC in combination with various detection systems, including UV and electrochemical detection (Cl7, C20, L23, L24, 04, S26), gas chromatography, gas chromatography-mass spectrometry (J2), electrospray mass spectrometry, and Western blotting or ELISA using antinitrotyrosine antibodies (H20, T2, V6). 

The effects of the experimental agents on lymphocyte activation and cytokine production were determined by ELISA using anticytokine antibodies. Testing results summarizing cytokine levels of tumor necrosis factor-a and interferon-y using nitro-polyunsaturated fatty chemical agents at 20 jlM are provided in Table 3. 

Nakahara, Y., Ishigami, A., Takeda, Y., Usagawa, T., and Uda, T., Enzyme Liidced Immunosorbent Assay (ELISA) using monoclonal antibody to detect methamphetamine in urine and hair, ]pn. /. Toxicol. Environ. Health, 35, 333,1989. 

To improve the specific response against microcystin immimoconjugates, synthetic lipopeptides were used as adjuvants and were found to invoke a greater immune response than the use of classical adjuvants such as Freunds adjuvant. However, an ELISA for the detection of free microcystin was not developed using these antibodies. Cross-reactivities of various microcystin variants and nodn-larin with monoclonal antibodies have been found affecting to the specificity of these antibodies for the recognition of the mentioned toxins. The number of purified microcystin variants that have been tested by ELISA using monoclonal antibodies shows marked differences between methods. 

The antigen then is mixed with serial dilutions of the enzyme-labeled antibody. A chromogenic substrate mixed with the conjugated enzyme yields a water-soluble product, the absorbency of which can be measured by a spectrophotometer. Recent technology has led to the development of rapid tests that do not require intact cells, live organisms, or cell cultures. ELISAs using monoclonal antibody techniques for the rapid detection of HSVs, adenoviruses, and C. trachomatis are highly sensitive and specific. 

RIA) using 2 I-labeled antibodies and enzyme-linked immunosorbent assay (ELISA) using alkaline phosphatase conjugates, wll be described. It is assumed that in most cases, the second antibodies will be bought either as purified material for radiolabeling or already conjugated to fluorescein, biotin, or the enzyme of choice (seeref. 2 for additional methods). 

Determination of the amount offosor mycprotein in the extract by ELISA using specific antibodies, one of which is conjugated to alkaline phosphatase (AP) and is detected by the AMPAK amplifier system. 

ELISA Using Enzyme-Labeled Antibody. Another type of competitive ELISA employs enzyme-labeled antibody with the antigen attached to a solid phase. In this technique, the binding of enzyme-labeled antibody to... 

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