UniProt Accession Number Reagent Type Target Name / Protein Biomarker Target Species Host Organism Isotype Clonality Vendor Catalog Number Conjugate RRID Availability Method Tissue Preservation Target Tissue Tissue State Detergent Antigen Retrieval Conditions Dye Inactivation Conditions Recommend Agree Disagree Contributor Notes
Q99JW5 Primary Antibody EpCAM Mouse Rat IgG2a G8.8 BioLegend 118222 AF594 AB_2563322 Stock IBEX2D Manual 1:4 Cytofix/Cytoperm Fixed Frozen Small Intestine NA 0.3% Triton-X-100 NA 1 mg/ml LiBH4 for more than 120 minutes Yes 0000-0003-4379-8967 [2] NA 0000-0003-4379-8967 1
P16422 Primary Antibody EpCAM Human Mouse IgG2b 9C4 BioLegend 324228 AF594 AB_2563209 Stock IBEX2D Manual 1:4 Cytofix/Cytoperm Fixed Frozen Lymph Node Metastatic 0.3% Triton-X-100 NA 1 mg/ml LiBH4 for more than 120 minutes Yes 0000-0003-4379-8967 [2, 1] NA 0000-0003-4379-8967 1
P16422 Primary Antibody EpCAM Human Mouse IgG2b 9C4 BioLegend 324228 AF594 AB_2563209 Stock Multiplexed 2D Imaging 1:4 Cytofix/Cytoperm Fixed Frozen Jejunum NA 0.3% Triton-X-100 NA NA Yes 0000-0003-4379-8967 NA 0000-0003-4379-8967  

Publications

  1. A. J. Radtke et al., “IBEX: an iterative immunolabeling and chemical bleaching method for high-content imaging of diverse tissues”, Nat. Protoc., 17(2):378-401, 2022, doi: 10.1038/s41596-021-00644-9.

    A. J. Radtke et al., “Accompanying dataset for: IBEX: An iterative immunolabeling and chemical bleaching method for high-content imaging of diverse tissues”, doi: 10.5281/zenodo.5244550.

  2. A. J. Radtke et al., “IBEX: A versatile multiplex optical imaging approach for deep phenotyping and spatial analysis of cells in complex tissues”, Proc Natl Acad Sci, 117(52):33455–33465, 2020, doi:10.1073/pnas.2018488117

Additional Notes

  1. Conjugate requires extended treatment with 1 mg/ml LiBH4 (120 minutes or more) to extinguish signal. Use in the last cycle or as a fiducial for IBEX experiments.
Mouse gut: EpCAM (magenta, catalog number 118222), CD8 (blue, catalog number 100738), and CD31 (green, catalog number 102516)
Mouse gut: EpCAM (blue, catalog number 118222), IgD (red, catalog number 405730), and CD11c (cyan, catalog number 117312)
Mouse gut: EpCAM (cyan, catalog number 118222), B220 (purple, catalog number 103248), and CD117 (yellow, catalog number 105827)
Mouse gut: EpCAM (blue, catalog number 118222), CD31 (magenta, catalog number 102516), and MHCII (green, catalog number 107622)
Human lymph node metastatic: CD4 (blue, catalog number 300526) and EpCAM (red, catalog number 324228)
Human jejunum: CD45 (cyan, catalog number 58-0459-41) and EpCAM (blue, catalog number 324228)