Name: Improving western blotting workflows
Uploaded: 2021-08-06T00:00:00.0000000
Duration: T00H02M25S
Description: Improving western blotting workflows

Name: Improving western blotting workflows

Description: Improving western blotting workflows

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Content URL: https://cadmoreoriginalmedia.blob.core.windows.net/c37b0ab7-9bd9-4f06-8fce-0273108850b1/western blotting IF V3 (2).mp4?sv=2019-02-02&sr=c&sig=TqjuEb%2FvJ39XPFaZLKGbm8cB7Et3yqIw2s7PeqFW1HE%3D&st=2024-11-21T17%3A51%3A30Z&se=2024-11-21T19%3A56%3A30Z&sp=r

Upload Date: 2021-08-06T00:00:00.0000000

Transcript: Language: EN.
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SPEAKER: Western blotting, or immunoblotting, has provided crucial information to characterize proteins in complex samples for over 40 years, and the technique is found in almost every biochemistry lab around the world due to its relatively cost-effective and simple nature. Although widespread, western blotting can be the source of fury and frustration, leveling experienced practitioners and hopeful novices alike with a seemingly random tendency to fail without much indication as to why.
SPEAKER: The western blotting technique is composed of five key steps, from sample preparation to data analysis, and so has numerous stages at which an error could occur. These stages include sample preparation, where sample cells are lysed to make the target proteins available in a suspension, unfolding the proteins and giving them a negative charge; protein separation, during which electrophoresis is used to separate the proteins by their mass charge ratio; blotting, which transfers the separated protein from the gel onto a blotting membrane; antibody incubation, at which stage a primary antibody binds the target protein, excess primary antibody is washed off, and a tagged secondary antibody is introduced that binds to the primary antibody, providing the readout for imaging; and finally, imaging and data analysis.
SPEAKER: Here, the reporter molecule of the secondary antibody is detected, establishing the presence and quantity of the target protein in the original sample. To take the frustration out of western blotting and to enable researchers to determine which stage an issue has arisen, new technologies are emerging. Such technologies include Bio-Rad Stain-Free western blotting. This technique removes the need to stain gels and membranes, allowing users to rapidly validate that key steps, such as electrophoresis and blotting, have been completed successfully.
SPEAKER: This avoids researchers applying expensive antibodies and detection reagents onto already failed blots, saving them time and money. [MUSIC PLAYING] To find out more about the modernization of western blotting and to discover how you could improve your western blotting workflows, check out our In Focus on western blotting, supported by Bio-Rad, over on www.biotechniques.com.

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