Dec 15, 2023

Public workspaceEa(52-68) uptake assays

Molecular Neurodegeneration
DOI
https://dx.doi.org/10.17504/protocols.io.14egn3r3pl5d/v1
  • Rebecca Wallings1
  • 1University of Florida
Rebecca Wallings
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DOI: https://dx.doi.org/10.17504/protocols.io.14egn3r3pl5d/v1
External link: https://doi.org/10.1186/s13024-025-00829-w
Protocol Citation: Rebecca Wallings 2023. Ea(52-68) uptake assays. protocols.io https://dx.doi.org/10.17504/protocols.io.14egn3r3pl5d/v1
Manuscript citation:
Wallings RL, Gillett DA, Staley HA, Mahn S, Mark J, Neighbarger N, Kordasiewicz H, Hirst WD, Tansey MG (2025) ASO-mediated knock-down of GPNMB in mutant-GRN and in Grn-deficient peripheral myeloid cells disrupts lysosomal function and immune responses. Molecular Neurodegeneration 20(). doi: 10.1186/s13024-025-00829-w
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
Keywords: uptake assays on pmac, uptake assay, pmac, uptake
Funders Acknowledgements:
ASAP
Grant ID: ASAP-020527
Abstract
Ea(52-68) uptake assays on pMacs
Troubleshooting
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