Jul 25, 2023
T cell depletion
- 1Columbia University
Protocol Citation: Connor Monahan 2023. T cell depletion. protocols.io https://dx.doi.org/10.17504/protocols.io.rm7vzx372gx1/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: July 13, 2023
Last Modified: May 31, 2024
Protocol Integer ID: 84950
Keywords: antibody, T cell, CD4 T cell, CD8 T cell, T cell depletion, flow cytometry, ASAPCRN
Funders Acknowledgement:
Aligning Science Across Parkinson's
Grant ID: 0375
Abstract
This protocol details T cell depletion by antibodies targeting CD4 and CD8.
Attachments
786-2014.pdf
37KB
Materials
Reagents
Ultra-LEAF™ Purified anti-mouse CD4 AntibodyBioLegendCatalog #100457
Ultra-LEAF™ Purified Rat IgG2b κ Isotype Ctrl AntibodyBioLegendCatalog #400644
Ultra-LEAF™ Purified anti-mouse CD8a AntibodyBioLegendCatalog #100746
Procedure
Procedure
Inject mouse intraperitoneally with 250 µg of Ultra-LEAF Purified anti-mouse CD4 or Rat IgG2b isotype control antibodies (Biolegend, Cat #100457 and Cat #400644; San Diego, CA) 3 and 1 days prior to immunization.
After immunization, inject mice with 250 µg of antibody weekly.
Assess cell depletion by flow cytometric analysis of mouse blood serum.
For depletion of CD8+ T cells, inject the mouse intraperitoneally with 250 µg of Ultra-LEAF Purified CD8+ or Rat IgG2b isotype control antibodies (Biolegend Cat # 100746 and Cat #400644; San Diego, CA) 2 days prior to immunization.
After immunization, inject mice with 250 µg of each antibody 9- and 19-days post immunization.
Assess cell depletion by flow cytometric analysis of mouse blood serum.