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
P16284 Primary Antibody CD31 Human Mouse IgG1 WM59 BioLegend 303106 PE AB_314332 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  
P16284 Primary Antibody CD31 Human Mouse IgG1 WM59 BioLegend 303106 PE AB_314332 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  
P16284 Primary Antibody CD31 Human Mouse IgG1 WM59 BioLegend 303106 PE AB_314332 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  
P16284 Primary Antibody CD31 Human Mouse IgG1 WM59 BioLegend 303106 PE AB_314332 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  
Q08481 Primary Antibody CD31 Mouse Rat IgG2a MEC13.3 BD Biosciences 553373 PE AB_394819 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] NA 0000-0003-4379-8967 1
Q08481 Primary Antibody CD31 Mouse Rat IgG2a MEC13.3 BD Biosciences 553373 PE AB_394819 Stock IBEX2D Manual 1:4 Cytofix/Cytoperm Fixed Frozen Thymus NA 0.3% Triton-X-100 NA 1 mg/ml LiBH4 15 minutes Yes 0000-0003-4379-8967 [2] NA 0000-0003-4379-8967  
Q08481 Primary Antibody CD31 Mouse Rat IgG2a MEC13.3 BD Biosciences 553373 PE AB_394819 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 [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., “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. Validated by Evelyn Kandov (No ORCID)
Human spleen: CD163 (yellow, based on catalog number 333602) and CD31 (blue, catalog number 303106)
Human lymph node: CD4 (cyan, catalog number 300520) and CD31 (magenta, catalog number 303106)
Human jejunum: Hoechst (blue, catalog number 40046), CD106 (cyan, catalog number 305806), and CD31 (red, catalog number 303106)