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
Q9NNX6 Primary Antibody DC-SIGN Human Mouse IgG2a 9E9A8 BioLegend 330112 AF647 AB_1186092 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  
Q9NNX6 Primary Antibody DC-SIGN Human Mouse IgG2b C209/1781 Novus Biologicals NBP2-54351AF647 AF647 NA Stock Multiplexed 2D Imaging FFPE Tonsil NA 0.3% Triton-X-100 pH 6 for 40 minutes at 95C (AR6 Akoya Biosciences AR600250ML) NA No 0000-0003-4379-8967 NA 0000-0003-4379-8967 1
Q9NNX6 Primary Antibody DC-SIGN Human Mouse IgG2b 102E11.06 Eurobio Scientific (Dendritics) DDX0202A647-100 AF647 NA Stock Multiplexed 2D Imaging FFPE Tonsil NA 0.3% Triton-X-100 pH 6 for 40 minutes at 95C (AR6 Akoya Biosciences AR600250ML) NA Yes 0000-0003-4379-8967 NA 0000-0003-4379-8967 2
Q9NNX6 Primary Antibody DC-SIGN Human Mouse IgG2a 9E9A8 BioLegend 330112 AF647 AB_1186092 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  
Q9NNX6 Primary Antibody DC-SIGN Human Mouse IgG2b 102E11.06 Eurobio Scientific (Dendritics) DDX0202A647-100 AF647 NA Stock Cell DIVE-IBEX 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 1 mg/ml LiBH4 15 minutes Yes 0000-0003-4379-8967 NA 0000-0003-4379-8967 3

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. Evaluated in human tonsil FFPE samples with HLA-DR (clone LN-3). Antibody does not label dendritic cells or macrophages. Recommend clones h209 or 102E11.06 for FFPE tissues and pH 6 antigen retrieval.
  2. Evaluated in human tonsil FFPE samples with HLA-DR (clone LN-3). Antibody labels dendritic cells and macrophages.
  3. Validated in human tonsil FFPE samples and shown to label dendritic cells and macrophage. Works beautifully on Cell DIVE imager with IBEX dye inactivation protocol.
Human lymph node: CD3 (purple, catalog number 300446) and DC-SIGN (cyan, catalog number 330112)
Human lymph node: DC-SIGN (magenta, catalog number 330112) and Vimentin (yellow, based on catalog number 677802)