Antibody anti-ferritin

To prepare an antibody protein array, a monolayer of protein A, which was compressed at a surface pressure of 11 mN m l was transferred to a compartment containing anti-ferritin antibody in 10 mM pH 7.0 phosphate buffer. The antibody molecules were self assembled onto the protein A layer. The protein A/antibody molecular membrane was transfered to a compartment containing ultrapure water for rinsing, and was then transfered onto the surface of an HOPG plate by the horizontal method. AFM measurements were made in a pH 7.0 of 10 mM phosphate buffer solution. 

One form of heterogeneous immunoassay is called enzyme-linked immunosorbent immunoassay (ELISA). In one instance, electrochemical immunoassay was performed for anti-ferritin (antibody) in a PDMS/PMMA chip. First, DTSSP was self-assembled on the gold electrode deposited on the PMMA plate. Then horse spleen ferritin (antigen) was attached to the DTSSP layer. A 100-p.g/mL solution of anti-horse ferritin (rabbit serum) was added. Then a secondary anti-rabbit antibody (HRP-linked) was introduced. A substrate (4-CN) was finally added which was converted to a precipitate product. The precipitate caused a reduction... 

The RIA-gnost Ferritin kit (no more commercially available) from formerly Behringwerke AG, Radiochemical Laboratory described below uses the principle of an immunoradiometric assay (IRMA). It is a two-site solid phase assay of the sandwich type, based on a plastic bead as solid phase to which the antiferritin antibody adheres. The antibody-solid phase is incubated with standards or serum samples containing ferritin and in this process the ferritin in the solution is bound quantitatively to the solid phase via the antibody. The amount of ferritin bound to the solid phase is then determined by a reaction with 125I-labeled anti-ferritin antibody. An antibody-ferritin-125I-antibody complex is thus formed. 

Dispense 300 pi of the anti-ferritin-125I-antibody solution into each tube and shake the test tubes rack briefly by hand. Then cover the rack with Parafilm and place it on the horizontal shaker and shake for 3 hours at room temperature (22 5 °C) with 300 rpm (250-350 rpm)... 

Pure PA is available commercially from Pharmacia Fine Chemicals, (Piscataway, New Jersey), who also produce PA bound covalently to Se-pharose 4B, and from Sigma Chemical Co. (St. Louis, Missouri). PA has been used mainly as an indicator of antibody bound to cell-surface anti-ggn. 14.15 por this purpose, it has been labeled with a fluorescent. tag (fluorescein 1 ), with ferritin, or with radionuclides. 1-, and H-labeled PA have been prepared, and their suitability as analytical reagents has been demonstrated. 

As with DNA above, a variety of proteins were immobilized non-covalently not only outside but also inside the CNT [138,150). Tsang et al. reported the first evidence for immobilization of small proteins inside MWNT by HRTEM [164-167], before DNA immobilization on the surface of MWNT as mentioned above [133,134]. After the innovative work by Sadler et aL, the cir-ciunference of CNTs was non-covalently functionalized with proteins directly or indirectly. MWNTs, prepared by arc discharge, were shown to be almost completely covered by streptavidin in helical arrangement [ 168]. SWNTs also adsorbed proteins and enzymes such as cytochrome c, ferritin, glucose oxidase and anti-fullerene IgG monoclonal antibody [169,170]. Indirect non-covalent functionaUzations of SWNTs with streptavidine and metallothionein were attained through triton-X 100-PEG-biotin and butylpyrene linkers, respectively [171,172]. 

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