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
P07766 Primary Antibody CD3 Human Mouse IgG1 UCHT1 BioLegend 300446 AF594 AB_2563236 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 [3, 1] NA 0000-0003-4379-8967 1
P22646 Primary Antibody CD3 Mouse Rat IgG2b 17A2 BioLegend 100240 AF594 AB_2563427 Stock IBEX2D Manual 1:4 Cytofix/Cytoperm Fixed Frozen Thymus NA 0.3% Triton-X-100 NA 1 mg/ml LiBH4 for more than 120 minutes Yes 0000-0003-4379-8967 [3] NA 0000-0003-4379-8967 1
P22646 Primary Antibody CD3 Mouse Rat IgG2b 17A2 BioLegend 100240 AF594 AB_2563427 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 [3] NA 0000-0003-4379-8967 1
P07766 Primary Antibody CD3 Human Mouse IgG1 UCHT1 BioLegend 300446 AF594 AB_2563236 Stock IBEX2D Manual 1:4 Cytofix/Cytoperm Fixed Frozen Lymph Node Follicular Lymphoma 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

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

  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.
Human lymph node: BCL2 (cyan, catalog number 658705) and CD3 (purple, catalog number 300446)
Human lymph node: CD3 (purple, catalog number 300446) and DC-SIGN (cyan, catalog number 330112)
Mouse thymus: CD3 (catalog number 100240)
Mouse lymph node: CD3 (cyan, catalog number 100240) and MHCII (blue, catalog number 107631)