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
P22646 Primary Antibody CD3 Mouse Rat IgG2b 17A2 BD Biosciences 557869 AF647 AB_396912 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
P22646 Primary Antibody CD3 Mouse Rat IgG2b 17A2 BD Biosciences 557869 AF647 AB_396912 Stock IBEX2D Manual 1:4 Cytofix/Cytoperm Fixed Frozen Small Intestine 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
P07766 Primary Antibody CD3 Human Mouse IgG1 UCHT1 BioLegend 300416 AF647 AB_389332 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, 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)
  2. Clone UCHT1 labels T cells in fixed frozen human tissues. Several conjugates are suitable for Multiplexed 2D Imaging; however, we recommend the following conjugates for IBEX experiments: AF488, AF555, PE, iF594, and AF647.