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The western blot is a widely used research technique that detects proteins using antibodies. To conduct a western blot, the proteins need to be transferred onto a membrane. There are different ways to transfer proteins onto the membrane. This video will be focusing on the wet transfer method.
The video lists the materials and reagents needed for a wet transfer and how to conduct it. Note that there are several different kinds of transfer buffers used. One of the most common is the Towbin transfer buffer, named after one of the scientists who helped develop the western blot. It contains 25mM of Tris, 19.2 mM of glycine and 20% methanol.
This video was created by Kieran Anderson, Kevin Cao, Aniqa Chaudhry, Brooke D’Mello, Ibrahim Ilahi, Kunwar Karim, Mubariz Maqsood, Adnan Moinuddin, Nardien Sedhom, and Julia Zhu in collaboration with the McMaster Demystifying Research Program.
We would like to also acknowledge the contributions of Dr. Takuma Isshiki, who conducted the wet transfer and provided guidance in the process. Additionally, we would like to also acknowledge Anmar Ayoub, who aided with materials and resources for the wet transfer. Finally, we would like to also thank Dr. Kjetil Ask and the Ask Lab for the additional guidance, insights and support.
Episode #1: Introduction to Western Blotting: • Introduction to the We...
Episode #2: SDS-PAGE: • Western Blot: Introduc...
Episode #4: Electroblotting: • Western Blot: Introduc...
Episode #5: Blocking: • Western Blot: Introduc...
Episode #6: Immunodetection: • Western Blot: Introduc...
Episode #7: Results & Analysis: • Western Blot: Result I...
This video is for general and educational purposes only.
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