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Sample preparation immobilization methods

Proteomics can be broadly defined as the identification, localization, and functional analysis of the protein make-up of cells. One of the major challenges faced, however, is lengthy sample preparation. Several methods have been developed to address this issue including the use of immobilized enzymes and acid-labile surfactants. Microwave heating can also play a valuable role. [Pg.248]

Fig. 11 Wet thickness (H) of PAAm in water as a function of the PAAm graft density for samples prepared by surface-initiated ATRP on substrates with gradient of initiator density. The initiator was immobilized by the vapor-diffusion method using mixtures of l-trichlorosilyl-2-(fn/p-chloromethyl phenyl)ethan n-octyl trichlorosilane (w/w) 1 1 (squares), 1 2 (circles), and 1 5 (triangles). The inset shows a cartoon illustrating the polymer behavior. Reproduced with permission from [134] (Copyright 2003 American Chemical Society)... Fig. 11 Wet thickness (H) of PAAm in water as a function of the PAAm graft density for samples prepared by surface-initiated ATRP on substrates with gradient of initiator density. The initiator was immobilized by the vapor-diffusion method using mixtures of l-trichlorosilyl-2-(fn/p-chloromethyl phenyl)ethan n-octyl trichlorosilane (w/w) 1 1 (squares), 1 2 (circles), and 1 5 (triangles). The inset shows a cartoon illustrating the polymer behavior. Reproduced with permission from [134] (Copyright 2003 American Chemical Society)...
HIAA. A ready-to-use competitive EIA kit has been developed for measuring chemically derivatized 5-HIAA. In this procedure, dichloromethane is used to convert 5-HIAA to its methyl ester during sample preparation. This methylated derivative competes with biotin-labeled 5-HIAA for a limited number of binding sites of an antibody immobilized on microtiter plates. After incubation, the wells are washed to remove unbound biotin-labeled HIAA and then incubated with antibiotin alkaline phosphatase before a p-nitrophenyl phosphate substrate is added. The amount of biotinylated HIAA bound to the antibody is inversely proportional to the 5-HIAA concentration. This EIA method shows excellent correlation with HPLC, and interassay reproducibility coefficient of variation (CV) is about 7% at a 5-HIAA level of 7.5mg/L. [Pg.1064]

The ICMs used for pesticide analysis include immimoassays (lAs) and the use of antibodies for sample preparation (e.g., for SPE and the cleanup of samples) [153], detection in flow-injection analysis, and biosensors. The earliest ICMs to be developed for pesticides analysis were lAs. There are various t) es of lAs, but the most frequently used in this context is the enzyme-linked immunosorbent assay (ELISA) [185]. ELISA is a heterogeneous assay because the antibodies or antigens are immobilized on a solid phase. Table 18.3 lists selected ELISA methods for the determination of pesticides in water samples [186-190]. Bjamason et al. have proposed an enzyme flow immunoassay (EFIA) using a protein G column for the determination of triazine herbicides in surface and wastewaters with a linear range between 0.1 and 10 pg/L [191]. [Pg.479]


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Immobilization methods

Sample methods

Sample preparation method

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