Western Blot Results

The quantitative Western blot data were confirmed using p53 and IκBα ELISA assays (Assay Designs, Ann Arbor, MI). The data produced with a Western blot is usually quite easy to interpret. In the majority of cases, bands corresponding to the target protein will become visible upon treatment of the blot with substrate.

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Western blot transfer apparatus.

Western blot results. The electrophoresis membrane is washed in a solution containing the primary antibody, before excess antibody is washed. However, the Western blot is no longer used, and today. 2.3.2 Results and discussion.

A high titer means, therefore, a high level of antibodies and is likely to indicate a major response by the body to a current or recent infection. Densitometric analysis of the Western blots indicated that the linear range for IκBα was 0.097–3.12 ng and for p53 was 1.87–60 ng. Schematic showing the assembly of a typical western blot apparatus with the position of the gel, transfer membrane, and direction of protein in relation to the electrode position.

Their identity is confirmed by comparison to molecular weight markers (for size) and a positive control (size and signal). The western blot, or western blotting, is a widely used analytical technique in molecular biology and immunogenetics to detect specific proteins in a sample of tissue homogenate or extract. This type of test is a serological assay for detecting antibodies to Borrelia bacteria that cause Lyme disease.

A synthetic or animal-derived antibody is created that recognises and binds to a specific target protein. The ELISAs were performed according to the manufacturer's instructions. In brief, the sample undergoes protein denaturation, followed by gel electrophoresis.

Results from a Western blot test are usually reported in terms of titers which is a measure of the amount of dilution a sample needs before no antibodies remain detectable. The result is a membrane with a copy of the protein pattern that was originally in the polyacrylamide gel. In case of a positive result from this test, the ELISA test was previously followed by a test called a Western blot to confirm the diagnosis.

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