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
P05362 Primary Antibody CD54 Human Mouse IgG1 HA58 BioLegend 353114 AF647 AB_2715942 Stock IBEX2D Automated 1:4 Cytofix/Cytoperm Fixed Frozen Jejunum NA 0.3% Triton-X-100 NA 0.5 mg/ml LiBH4 10 minutes continuous exchange with automated protocol Yes 0000-0003-4379-8967 [1] NA 0000-0003-4379-8967  
P05362 Primary Antibody CD54 Human Mouse IgG1 HA58 BioLegend 353114 AF647 AB_2715942 Stock IBEX2D Manual 1:4 Cytofix/Cytoperm Fixed Frozen Liver NA 0.3% Triton-X-100 NA 1 mg/ml LiBH4 15 minutes Yes 0000-0003-4379-8967 [2, 1] NA 0000-0003-4379-8967  
P05362 Primary Antibody CD54 Human Mouse IgG1 HA58 BioLegend 353114 AF647 AB_2715942 Stock IBEX2D Automated 1:4 Cytofix/Cytoperm Fixed Frozen Lymph Node NA 0.3% Triton-X-100 NA 0.5 mg/ml LiBH4 10 minutes continuous exchange with automated protocol Yes 0000-0003-4379-8967 [1] NA 0000-0003-4379-8967  
P05362 Primary Antibody CD54 Human Mouse IgG1 HA58 BioLegend 353114 AF647 AB_2715942 Stock IBEX2D Automated 1:4 Cytofix/Cytoperm Fixed Frozen Skin NA 0.3% Triton-X-100 NA 0.5 mg/ml LiBH4 10 minutes continuous exchange with automated protocol Yes 0000-0003-4379-8967 [1] NA 0000-0003-4379-8967  
P05362 Primary Antibody CD54 Human Mouse IgG1 HA58 BioLegend 353114 AF647 AB_2715942 Stock IBEX2D Manual 1:4 Cytofix/Cytoperm Fixed Frozen Spleen NA 0.3% Triton-X-100 NA 1 mg/ml LiBH4 15 minutes Yes 0000-0003-4379-8967 [2, 1] 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

Human spleen: CD21 (red, based on catalog number 354902) and CD54 (blue, catalog number 353114)
Human liver: CD54 (cyan, catalog number 353114) and CD138 (purple, catalog number 356504)
Human jejunum: Hoechst (blue, catalog number 40046), CD54 (green, catalog number 353114), and Vimentin (magenta, catalog number 677807)