immunoblots

본 내용은 " immunoblots " 의 원문 자료에서 일부 인용된 것입니다. 2024.09.13

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• 1. Immunoblotting

  Immunoblotting(western blotting)은 전기영동 후 단백질, 특히 양이 적은 단백질의 면역검출을 위한 강력하고 중요한 절차이다. Immunoblotting을 통해 nitrocellulose 또는 polyvinylidene fluoride(PVDF)와 같은 단백질 유지 막 지지체에 고정된 단백질 항원을 검출할 수 있다. 관심 단백질의 검출은 membrane에 노출된 관심 단백질을 특이적으로 인식하는 항체의 결합에 의존한다.

• 2. House keeping gene

  House keeping gene은 대부분의 세포 유형 및 다양한 실험 조건 하에서 비교적 일정한 수준으로 상수적으로 발현되는 유전자이다. House keeping gene은 단백질 로딩을 위한 내부 대조군과 western blott 분석의 참조로 사용된다. 가장 일반적으로 사용되는 하우스키핑 유전자는 β-actin, β-tubulin, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH)이다.

• 3. Antibody detection

  Western blot은 단백질을 분리하고 식별하기 위한 연구에서 종종 사용된다. 이 기술에서는 겔 전기영동을 통해 단백질 혼합물을 분자량에 따라 유형별로 분리한다. 그런 다음 이 결과는 각 단백질에 대한 밴드를 생성하는 막으로 전달된다. 그런 다음 멤브레인을 관심 단백질에 특이적인 라벨 항체와 함께 배양한다. 결합되지 않은 항체는 씻어내고 관심 단백질에 결합된 항체만 남긴다. 결합된 항체는 필름을 현상하여 검출된다.

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• 1. Immunoblotting

  Immunoblotting, also known as 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 identify and visualize the target proteins. This method is widely used in various fields, including molecular biology, biochemistry, and clinical diagnostics, to study protein expression, post-translational modifications, and protein-protein interactions. Immunoblotting is a highly sensitive and specific technique that allows researchers to gain valuable insights into the biological processes and pathways within cells and tissues. Its ability to provide both qualitative and quantitative information about proteins makes it an indispensable tool in modern scientific research.

• 2. House keeping gene

  Housekeeping genes, also known as reference genes, are genes that are constitutively expressed in all cells and are essential for the basic cellular functions necessary for cell survival and proliferation. These genes are often used as internal controls in gene expression studies, such as quantitative real-time PCR (qRT-PCR) and RNA-sequencing, to normalize the expression levels of target genes. The rationale behind using housekeeping genes as references is that their expression levels should remain relatively constant across different experimental conditions, cell types, and developmental stages. However, it is important to note that the expression of even so-called 'stable' housekeeping genes can vary under certain circumstances, and the selection of appropriate reference genes should be carefully evaluated for each specific experimental setup. The appropriate use of housekeeping genes is crucial for the accurate interpretation of gene expression data and the reliable quantification of target gene expression.

• 3. Antibody detection

  Antibody detection is a fundamental technique in immunology and diagnostic medicine, used to identify the presence and levels of specific antibodies in biological samples. This is crucial for the diagnosis and monitoring of various infectious diseases, autoimmune disorders, and other conditions where the immune system plays a central role. Commonly used methods for antibody detection include enzyme-linked immunosorbent assay (ELISA), immunofluorescence assay, and Western blotting. These techniques rely on the specific binding of antibodies to their target antigens, allowing for the sensitive and quantitative measurement of antibody levels. Accurate antibody detection is essential for disease surveillance, vaccine development, and understanding the dynamics of the immune response. As our understanding of the immune system continues to evolve, the development of more advanced and sensitive antibody detection methods will be crucial for advancing both clinical and research applications in the field of immunology.