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
P01732 Primary Antibody CD8 Human Mouse IgG1 SK1 BioLegend 344716 AF488 AB_10549301 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  
P01732 Primary Antibody CD8 Human Mouse IgG1 SK1 BioLegend 344716 AF488 AB_10549301 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 [3, 1] NA 0000-0003-4379-8967  
P01732 Primary Antibody CD8 Human Mouse IgG1 SK1 BioLegend 344716 AF488 AB_10549301 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  
P01732 Primary Antibody CD8 Human Mouse IgG1 SK1 BioLegend 344716 AF488 AB_10549301 Stock IBEX2D Manual 1:4 Cytofix/Cytoperm Fixed Frozen Lymph Node NA 0.3% Triton-X-100 NA 1 mg/ml LiBH4 15 minutes Yes 0000-0003-4379-8967 [3, 1] NA 0000-0003-4379-8967  
P01732 Primary Antibody CD8 Human Mouse IgG1 SK1 BioLegend 344716 AF488 AB_10549301 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 [3, 1] NA 0000-0003-4379-8967  
P01731 Primary Antibody CD8 Mouse Rat IgG2a 53-6.7 BioLegend 100723 AF488 AB_389304 Stock IBEX2D Manual 1:4 Cytofix/Cytoperm Fixed Frozen Lymph Node NA 0.3% Triton-X-100 NA 1 mg/ml LiBH4 15 minutes Yes 0000-0003-4379-8967 [3] NA 0000-0003-4379-8967  
P01732 Primary Antibody CD8 Human Mouse IgG1 SK1 BioLegend 344716 AF488 AB_10549301 Stock IBEX2D Manual 1:4 Cytofix/Cytoperm Fixed Frozen Lymph Node Follicular Lymphoma 0.3% Triton-X-100 NA 1 mg/ml LiBH4 15 minutes Yes 0000-0003-4379-8967 [2] 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., “A Multi-scale, Multiomic Atlas of Human Normal and Follicular Lymphoma Lymph Nodes”, bioRxiv, 2022, doi: 10.1101/2022.06.03.494716.

  3. 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 lymph node: CD1c (red, catalog number 331506), CD8 (blue, catalog number 344716), and CD25 (yellow, catalog number 356128)
Human lymph node metastatic: CD8 (green, catalog number 344716) and CD138 (blue, catalog number 356524)
Human lymph node: CD4 (cyan, catalog number 58-0049-42) and CD8 (red, catalog number 561525)
Human spleen: CD8 (cyan, catalog number 344716) and CD3 (red, catalog number 1053135)