- The western blot method is a protein detection technique that involves the following steps1234:
- Sample preparation: Proteins are extracted from different samples, such as tissues or cells.
- Gel electrophoresis: The most commonly used gel is polyacrylamide gels (PAG) and buffers loaded with sodium dodecyl sulfate (SDS).
- Proteins transfer: The separated molecules are transferred or blotted onto a second matrix, generally a nitrocellulose or polyvinylidene difluoride (PVDF) membrane.
- Blocking: The membrane is blocked to prevent non-specific binding of the primary antibody.
- Treatment with Primary Antibody: The membrane is incubated with a primary antibody that binds to the protein of interest.
- Treatment with secondary antibody: The membrane is incubated with a secondary antibody that binds to the primary antibody.
- Treatment with suitable substrate: The substrate reacts with the enzyme conjugated to the secondary antibody and produces a detectable signal.
了解详细信息：✕此摘要是使用基于多个在线源的 AI 生成的。若要查看原始源信息，请使用“了解详细信息”链接。The western blot method is composed of a gel electrophoresis to separate native proteins by 3-D structure or denatured proteins by the length of the polypeptide, followed by an electrophoretic transfer onto a...
en.wikipedia.org/wiki/Western_blotThe Principle and Procedure of Western Blot
www.creative-proteomics.com/blog/index.php/the-p…Procedure/Steps:
www.onlinebiologynotes.com/western-blotting-tech…The first step in a western blotting procedure is to separate the macromolecules in a sample using gel electrophoresis. Subsequently, the separated molecules are transferred or blotted onto a second matrix,...
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Learn how to perform western blotting with this comprehensive guide from Abcam. Find solutions and reagents, procedures, and useful links for sample lysis, gel electrophoresis, transfer, and antibody staining. 展开
- Preparation of lysate from cell culture
- 1. Place the cell culture dish on ice and wash the cells with ice-cold PBS. 2. Aspi…
- Preparation of lysate from tissues
- 1. Dissect the tissue of interest with clean tools, on ice preferably, an… 展开
The membrane can be either nitrocellulose or PVDF. Activate PVDF with methanol for 1 min and rinse with transfer buffer before preparing the stack. The time and voltage of transfer may require … 展开
- 1. Remove a small volume of lysate to perform a protein quantification assay. D…
- 2. Determine how much protein to load and add an equal volume 2X Laemmli sampl…
- 3. To reduce and denature your samples, boil each cell lysate in sam… 展开
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网页Learn the basics of western blotting, a technique for protein analysis using antibodies and gels. Find out how to transfer, block, probe and detect proteins on a membrane.
网页2023年5月28日 · Learn how to perform western blot, a technique to separate and identify proteins in a complex sample. Follow the step-by-step procedure, see the result interpretation, and explore the …
Western Blot: Technique, Theory, and Trouble Shooting - PMC
网页Western blotting is an important technique used in cell and molecular biology. By using a western blot, researchers are able to identify specific proteins from a complex mixture of …
网页Learn how to perform western blotting, a technique for detecting and analyzing proteins, in seven basic steps. Find out about sample preparation, gel electrophoresis, transfer, …
Western Blotting: An Introduction - PMC - National Center for ...
网页Western blotting is an important procedure for the immunodetection of proteins, particularly proteins that are of low abundance. This process involves the transfer of protein patterns …
Western blot protocol | abcam
网页Western blotting is a technique that uses specific antibodies to identify proteins separated by size through gel electrophoresis. The immunoassay uses a membrane made of …
网页Western Blot is a method for detecting the presence of specific proteins from mixture of proteins. Sources. Western Blotting Technique in Biomedical Research. Using Western …
网页Western blotting (WB) is widely used to analyze specific protein expression in cell or tissue extracts. At Cell Signaling Technology (CST) we understand that western blotting …
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