Nov 23, 2023
iPSC-derived neurons sample preparation for ELISA assays
- 1The Francis Crick Institute, 1 Midland Road, London NW1 1AT;
- 2Department of Clinical and Movement Neurosciences, UCL Queen Square Institute of Neurology, Queen Square, London, WC1N 3BG
Protocol Citation: Laura Mahoney-Sanchez 2023. iPSC-derived neurons sample preparation for ELISA assays. protocols.io https://dx.doi.org/10.17504/protocols.io.dm6gp35jdvzp/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 07, 2023
Last Modified: November 23, 2023
Protocol Integer ID: 90561
Abstract
Cell lysis sample preparation for ELISA assays
Cell lysis
Cell lysis
Cells were washed with cold PBS then lysed with 300 µL cold RIPA buffer (Thermo Fisher Scientific) supplemented with 1x of a cocktail of phosphatase and protease inhibitors (Life Technologies).
Cell lysates were transferred into 1.5ml eppendorf tubes and centrifuged for 00:05:00 at 3500rpm to remove cell debris.
5m
Following centrifugation, the supernatant was collected and protein concentration were measured using a BCA or Bradford protein assay (Thermo Fisher Scientific).
ELISA assay
ELISA assay
Equal amounts of protein lysate, 20 µg , were used to perform ELISA assays according to manufacturer's instructions.