Proteins combine with membrane based on a hydrophobic interaction, thereby having a slight effect on protein activities. Proteins move within the gel onto a membrane made of nitrocellulose (NC) or polyvinylidene difluoride (PVDF). Western blotting is based on the principle of immunochromatography where proteins are separated onto polyacrylamide gel according to their molecular weight. The western blot is also used for medical diagnoses such as HIV-test and BSE-test. Besides, a dilution series of a purified protein of known concentrations can be used for precise estimation of protein concentration. Semi-quantitative estimation of a protein can also be derived from the color intensity and the size of the band on the blot membrane. It is generally used as an analytical method to identify the presence of a specific single protein within a complex mixture. The western blot is widely used in biochemistry for the qualitative evaluation of single proteins and protein-modifications (such as post-translational modifications). The western blot is a powerful analytical tool used in molecular biology, immunogenetics and other molecular biology disciplines for protein characterization from a sample of tissue homogenate or extract. Western blotting facilitates the qualitative and quantitative assessment of protein expression in a variety of tissues and allows inferences about pre- and post-translational processes (e.g., phosphorylation or alternative splicing). The membrane containing bands is then incubated with antibodies specific to the target protein. The results of the electrophoresis are then transferred to a polyvinylidene fluoride (PVDF) membrane producing a band for each protein. In this technique, the proteins are assorted based on their molecular weight, and type, through gel electrophoresis. The Western blot is a biological technique that allows for the specific identification and characterization of proteins. Balances, Scales and Weighing Equipment.
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