Monoclonal Antibody Characterization Protocol

The mouse anti-HA monoclonal antibody (mAb) and the mouse anti-human IgG (29.5; Fc specific; anti-Fc) were generated in our lab and have been previously described (17 (link)). The anti-human IgG (Fc specific) labelled with peroxidase (POD) was from Sigma-Aldrich, the anti-human PD-1 (clone EH12.2H7) from BioLegend, and the anti-mouse IgG-PE and IgG-APC from Jackson ImmunoResearch. The rabbit anti-HA (clone C29F4) and the anti-β-actin (clone C4) mAbs, and the anti-rabbit IgG-POD, and the anti-mouse IgG-HRP, were from Cell Signaling, MP Biomedicals, Promega, and Sigma-Aldrich, respectively. The streptavidin-AF555 and the anti-rabbit IgG (H+L)-biotinylated were from Life Technologies and Jackson Immunoresearch, respectively. The anti-human CD14-APC (clone M5E2) mAb employed to stain surface expression of anti-CD3 single chain fragments, and the anti-mouse CD45-APC (clone 104) mAb used to exclude the TCS in the reporter assays, were obtained from BioLegend.

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Martínez-Vicente P., Poblador F., Leitner J., Farré D., Steinberger P., Engel P, & Angulo A. (2022). Discovery of the first PD-1 ligand encoded by a pathogen. Frontiers in Immunology, 13, 1007334.

Publication 2022

anti-human PD-1 (clone EH12.2H7) rabbit anti-HA (clone C29F4) anti-mouse IgG-HRP

Actin Anti antibody Anti cd3 Anti igg Assays Clone Human Monoclonal antibody Mouse Peroxidase Promega Rabbit Stain Streptavidin

Corresponding Organization : Consorci Institut D'Investigacions Biomediques August Pi I Sunyer

Other organizations : Medical University of Vienna

Top 5 similar protocols

Variable analysis

independent variables

Mouse anti-HA monoclonal antibody (mAb)
Mouse anti-human IgG (29.5; Fc specific; anti-Fc)
Anti-human IgG (Fc specific) labelled with peroxidase (POD)
Anti-human PD-1 (clone EH12.2H7)
Anti-mouse IgG-PE and IgG-APC
Rabbit anti-HA (clone C29F4)
Anti-β-actin (clone C4) mAbs
Anti-rabbit IgG-POD
Anti-mouse IgG-HRP
Streptavidin-AF555
Anti-rabbit IgG (H+L)-biotinylated
Anti-human CD14-APC (clone M5E2) mAb
Anti-mouse CD45-APC (clone 104) mAb

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

positive controls

Not specified

negative controls

Not specified

Annotations

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