Samples were analysed using an ELISA designed to detect and measure the level of α-synuclein protein in cell lysates. ELISA stands for Enzyme Linked-Immuno-Sorbent Assay. It is a very sensitive and specific assay and is used in health research and biochemistry to detect if a certain substance is present in a sample. An ELISA assay uses antibodies, specific to the susbtance, that are linked to an enzyme. The enzyme causes chromogen to produce a fluorescent signal, whose magnitude depends on the amount of the susbtance present.
Using a standard curve, the readings from the ELISA reader were interpreted and the concentration of alpha-synuclein in each of the samples was calculated.
BCA Protein Assay Results
The BCA assay, also known as the Microparticle-bound Protein Assay is a simple yet accurate way of measuring the total amount of protein in a sample. This direct measurement is possible using the copper reduction/bicinchoninic acid (BCA) reaction. Copper (II) is reduced to copper (I) by protein under alkaline conditions. The copper (I) ion generated reacts with the BCA to form a purple solution whose fluorescent signal depends on the total amount of protein in the assay.
Using a standard curve, the readings from the ELISA reader were interpreted and the concentration of total protein in each of the samples was calculated.
Determining the amount of α-synuclein relative to the amount of total protein
After determining the concentrations of alpha-synuclein and of total protein, the amount of α-synuclein relative to the amount of total protein was calculated. It was determined through dividing the average concentration of α-synuclein by the average concentration of total protein. This ensured that the results were accurate and that only the amount of alpha-synuclein had been measured.
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