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
Q08481 Primary Antibody CD31 Mouse Rat IgG2a MEC13.3 BioLegend 102520 AF594 AB_2563319 Stock IBEX2D Manual 1:4 Cytofix/Cytoperm Fixed Frozen Lung 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
P16284 Primary Antibody CD31 Human Mouse IgG1 WM59 BioLegend 303126 AF594 AB_2563303 Stock IBEX2D Manual 1:4 Cytofix/Cytoperm Fixed Frozen Lymph Node NA 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 2

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). 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.
  2. Clone WM59 labels endothelial cells in various tissues and works in several formats (AF488, AF532, AF647, AF700). AF594 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 lung: Ly-6G (yellow, catalog number 127626), CD31 (blue, catalog number 102520), and CD8 (red, catalog number 100724)