Electrophoresis is a widely used technique in molecular biology and biochemistry to separate and analyze proteins based on their size, charge, and other physical properties. The composition of a protein sample can be determined using various types of electrophoresis techniques, which are briefly discussed below:1. Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis SDS-PAGE : SDS-PAGE is the most common electrophoresis technique used for protein separation. In this method, proteins are denatured and coated with the negatively charged detergent SDS, which imparts a uniform charge-to-mass ratio. The proteins are then separated based on their molecular weight in a polyacrylamide gel matrix under an electric field. Smaller proteins migrate faster through the gel, while larger proteins move slower. After electrophoresis, the gel can be stained with a protein-specific dye, and the bands can be visualized and analyzed.2. Native-PAGE: Unlike SDS-PAGE, native-PAGE does not use SDS to denature proteins. Instead, proteins are separated based on their native charge and size. This technique is useful for studying protein-protein interactions, enzyme activity, and protein conformation.3. Isoelectric Focusing IEF : IEF is a technique used to separate proteins based on their isoelectric point pI , which is the pH at which a protein has no net charge. In this method, a pH gradient is established in a gel, and proteins are separated according to their pI values. Proteins with a pI lower than the pH of the gel will migrate towards the anode positive electrode , while proteins with a pI higher than the pH of the gel will migrate towards the cathode negative electrode .4. Two-Dimensional Gel Electrophoresis 2D-PAGE : This technique combines IEF and SDS-PAGE to separate proteins based on both their pI and molecular weight. In the first dimension, proteins are separated by IEF, and in the second dimension, they are separated by SDS-PAGE. This method provides high-resolution separation and is useful for analyzing complex protein mixtures, such as cellular lysates.5. Capillary Electrophoresis CE : CE is a high-resolution separation technique that uses narrow capillaries filled with a separation medium e.g., polyacrylamide gel instead of a slab gel. Proteins are separated based on their size and charge under an electric field. CE is highly sensitive and requires a small sample volume, making it suitable for analyzing trace amounts of proteins.Real-life application of electrophoresis in protein analysis:One real-life application of electrophoresis in protein analysis is in the diagnosis of genetic diseases caused by abnormal protein production or mutations. For example, hemoglobin electrophoresis is used to diagnose hemoglobinopathies, such as sickle cell anemia and thalassemia. In this technique, hemoglobin proteins are separated based on their charge and size, allowing for the identification of abnormal hemoglobin variants. Early diagnosis of these diseases can help in the management and treatment of the affected individuals.