Dec 15, 2023
Ea(52-68) uptake assays
- 1University of Florida
Protocol Citation: Rebecca Wallings 2023. Ea(52-68) uptake assays. protocols.io https://dx.doi.org/10.17504/protocols.io.14egn3r3pl5d/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: December 14, 2023
Last Modified: December 15, 2023
Protocol Integer ID: 92327
Funders Acknowledgement:
ASAP
Grant ID: ASAP-020527
Abstract
Ea(52-68) uptake assays on pMacs
Reconstitute MHC-II Ea chain (Ea) (52–68) peptide (Anaspec) in sterile distilled H2O to a final concentrate of 1mg/mL.
Once peritoneal macrophages are adhered to plates, add 5µg per well of 24-well plate and incubate for 18-hours
Harvest cells as per flow cytometry protocol and stain with eBioY-Ae antibody (ThermoFisher) to assess Ea52-68 peptide bound to I-Ab