License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it’s working
Created: March 04, 2022
Last Modified: March 04, 2022
Protocol Integer ID: 59052
Abstract
This protocol is used to western blot proteins-of-interest.
Add 4X Laemmli buffer to protein samples and incubate at 95oC for 7 minutes.
7m
Load samples into 12% polyacrylamide gel and run in running buffer at 80-100V for 60-120 minutes.
2h
Transfer gel to 0.2μm nitrocellulose membrane at 350mA for 60 minutes at 4oC.
1h
Wash membrane with 1X TBS-T then block in 10% milk for 30 minutes at room temperature.