Nov 12, 2023
Pull-down Assays
- 1University of California, San Diego
Protocol Citation: Leonardo A Parra-Rivas 2023. Pull-down Assays. protocols.io https://dx.doi.org/10.17504/protocols.io.x54v9pw5pg3e/v1
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: November 12, 2023
Last Modified: November 12, 2023
Protocol Integer ID: 90822
Abstract
Pull-down Assays
To pull down proteins from brain lysates and LP2(P4) fractions, 1-2 mg of the
sample was incubated with 25-50 μg of glutathione beads containing GST fusion
proteins for 12-16 hrs.
The mixtures were washed three times with 1X PBS with
0.15% Triton X-100, and then resuspended in 20 μl of 1X NuPAGE LDS sample
buffer for NuPAGE and immunoblotted analysis.
Alternatively, the samples were
resuspended in 50μl of PBS for proteomics studies and stored at -20 °C before
quality control and proteomics analysis.