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
P01903 Primary Antibody HLA-DR Human Mouse IgG2a L243 BioLegend 307620 AF488 AB_493175 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  
P01903 Primary Antibody HLA-DR Human Mouse IgG2a L243 BioLegend 307620 AF488 AB_493175 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  
P01903 Primary Antibody HLA-DR Human Mouse IgG2a L243 BioLegend 307620 AF488 AB_493175 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  
P01903 Primary Antibody HLA-DR Human Mouse IgG2a L243 BioLegend 307619 AF488 AB_493176 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  
P01903 Primary Antibody HLA-DR Human Mouse IgG2a L243 BioLegend 307620 AF488 AB_493175 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  
P01903 Primary Antibody HLA-DR Human Mouse IgG2b LN-3 Novus Biologicals NBP2-47670AF488 AF488 NA Stock Multiplexed 2D Imaging FFPE Tonsil NA 0.3% Triton-X-100 pH 6 for 30 minutes ER1 (AR9961) and pH 9 for 30 minutes ER2 (AR9640) using the Leica Bond NA Yes 0000-0003-4379-8967 NA 0000-0003-4379-8967 1
P01903 Primary Antibody HLA-DR Human Mouse IgG2b LN-3 Novus Biologicals NBP2-47670AF488 AF488 NA Stock Multiplexed 2D Imaging FFPE Lymph node Follicular Lymphoma 0.3% Triton-X-100 pH 6 for 30 minutes ER1 (AR9961) and pH 9 for 30 minutes ER2 (AR9640) using the Leica Bond NA Yes 0000-0003-4379-8967 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. Labels macrophages, dendritic cells, and B cells in FFPE tissues.
Human lymph node: HLA-DR (cyan, catalog number 307620) and CD69 (red, catalog number 310918)
Human lymph node: HLA-DR (purple, catalog number 307620) and Valpha7.2 (yellow, catalog number 351726)
Human liver: HLA-DR (cyan, catalog number 307620) and Vimentin (red, catalog number 677807)
Human FFPE follicular lymphoma lymph node: HLA-DR (red, catalog number NBP2-47670AF488)
Human FFPE follicular lymphoma lymph node: HLA-DR (red, catalog number NBP2-47670AF488) and PD-1 (cyan, catalog number ab275126)
Human lymph node: HLA-DR (green, catalog number 307620)