Serum solution preparation

Dilute the various rabbit serum antibody solutions prepared on Day 1 in hlocking buffer as follows ... 

Atomic Absorption Assay for Copper in Serum Standard Copper Solutions. Prepare a solution of copper containing 1 mg in 1 ml as described in die colorimetric method, above. Serially dilute the solution widi sodium chloride-potassium chloride solution to produce solutions containing 0.5, 1.0,... 

Plasmid DNA solution prepare DNA solution e.g. luciferase reporter plasmid or eGFP plasmid, at a concentration of 12 pg DNA/ml by dilution of the stock solution with a serum- and supplement-free medium (e.g., RPMl 1640). 

In this section, the application of the sensing mechanism described in Section 3.1 to an in vitro environmental model containing interfering cell culture medium is described. In this experiment the cell culture medium was used in place of deionized water, that is, 0.5 ml DMEM F-12 culmre media containing 1% fetal bovine serum (FBS) was mixed with 5.5 ml AuNPs stock solution to obtain a final AuNPs concentration of 2.25 X 10 NPs ml with pH 6.85. Additionally, to adjust the ion concentration, as was done previously, 100 pi of 10 mM FeCl3 at pH 2.90 was added into the mixture. Finally, 1 ml of 15 mM GSH at pH 3.44 was added to mediate the AuNPs. All optical characterizations were performed within 6 h of solution preparations. 

Calibrate using cystine solutions prepared by dissolving it in IN NH solution in concentrations of 2, 4, 6, 8 and 10 mg/100 ml, polarographing them in the same volumes as urine or serum appropriately diluted. Such a calibration, however, is not sufiicient for the determination of cystine in hydrolysates in which the presence of some other amino acids (e.g.,... 

Incubate sections/cells with a blocking solution prepared with NGS and NDS (10 % each) in PBS (use always normal serum from the same species in which secondary antibodies were raised) or 1 % BSA in PBS for 30 min at RT see Note 6). In parallel, run one to two samples as negative controls, where the blocking solution will be applied to sections/cells instead of the primary antibodies, under the same time and temperature conditions see Note 7). 

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