Western Blotting

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1. Western Blotting

Western blotting은 sample 내 특정 단백질의 존재 여부나 크기, 양 등을 확인하는 실험법으로, 해당 단백질과 결합하는 antibody를 이용하여 단백질을 찾아내는 방식을 이용한다. 이 실험을 통해 western blotting의 과정과 원리를 이해할 수 있다.
2. SDS-PAGE

SDS-PAGE는 SDS(Sodium Dodecyl Sulfate)로 coating 된 단백질을 2PAGE(Polyacrylamide Gel Electrophoresis)를 통해 크기에 따라 분리하는 실험 기법이다. 이를 통해 더욱 정밀한 단백질 존재 여부 파악이 가능하다.
3. Antibody

Antibody는 B cell에서 생산되는 γ-면역 글로불린 당단백질로, antigen의 epitope와 specific한 결합을 할 수 있는 성질이 있어 western blotting 실험 시 주로 사용된다. 1차 antibody와 2차 antibody를 사용하여 단백질을 detection 한다.

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1. Western Blotting

Western blotting is a powerful analytical technique used to detect and quantify specific proteins in a complex mixture. It involves separating proteins by size using gel electrophoresis, transferring them to a membrane, and then using antibodies to detect and visualize the target proteins. This method is widely used in various fields of biology and medicine, as it allows researchers to study protein expression, modifications, and interactions. Western blotting is a versatile and sensitive technique that provides valuable insights into the molecular mechanisms underlying biological processes. It is an essential tool for understanding protein function, disease pathogenesis, and the development of targeted therapies.
2. SDS-PAGE

SDS-PAGE (Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis) is a widely used technique in molecular biology and biochemistry for the separation and analysis of proteins based on their molecular weight. This method involves the denaturation of proteins using the anionic detergent SDS, which binds to the proteins and gives them a uniform negative charge. The proteins are then separated through a polyacrylamide gel matrix under an electric field, with smaller proteins migrating faster than larger ones. SDS-PAGE is a crucial tool for protein characterization, purification, and identification, and is often used in conjunction with other techniques, such as Western blotting, to provide a comprehensive understanding of the protein composition and structure within a sample. Its simplicity, reproducibility, and ability to resolve complex protein mixtures make SDS-PAGE an indispensable technique in modern biochemistry and molecular biology research.
3. Antibody

Antibodies are remarkable biomolecules that play a crucial role in the immune system's defense against foreign pathogens and substances. These Y-shaped proteins are produced by specialized immune cells called B lymphocytes and are capable of recognizing and binding to specific target molecules, known as antigens. Antibodies can neutralize pathogens, mark them for destruction by other immune cells, and facilitate the clearance of harmful substances from the body. The ability to generate and utilize antibodies has revolutionized various fields, including diagnostics, therapeutics, and research. Monoclonal antibodies, in particular, have become invaluable tools for targeted therapies, while polyclonal antibodies are widely used in immunoassays and immunohistochemistry. The continued advancement in antibody engineering and production has expanded the applications of these versatile biomolecules, making them indispensable in modern medicine, biotechnology, and scientific investigation.

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