Nov 13, 2023

Pull-down Assays V.1

DOI
https://dx.doi.org/10.17504/protocols.io.x54v9pw5pg3e/v1
  • 1University of California, San Diego
Leonardo A Parra-Rivas
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DOI: https://dx.doi.org/10.17504/protocols.io.x54v9pw5pg3e/v1
Protocol Citation: Leonardo 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 13, 2023
Last Modified: November 13, 2023
Protocol  Integer ID: 90822
Keywords: assay
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.