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
P08670 Primary Antibody Vimentin Human Mouse IgG2a O91D3 BioLegend 677807 AF647 AB_2616801 Stock IBEX2D Automated 1:4 Cytofix/Cytoperm Fixed Frozen Jejunum 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  
P08670 Primary Antibody Vimentin Human Mouse IgG2a O91D3 BioLegend 677807 AF647 AB_2616801 Stock IBEX2D Automated FFPE Kidney NA 0.3% Triton-X-100 pH 6 for 40 minutes at 95C (AR6 Akoya Biosciences AR600250ML) 0.5 mg/ml LiBH4 10 minutes continuous exchange with automated protocol Yes 0000-0003-4379-8967 [1] NA 0000-0003-4379-8967  
P08670 Primary Antibody Vimentin Human Mouse IgG2a O91D3 BioLegend 677807 AF647 AB_2616801 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  
P08670 Primary Antibody Vimentin Human Mouse IgG2a O91D3 BioLegend 677807 AF647 AB_2616801 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  
P08670 Primary Antibody Vimentin Human Mouse IgG2a O91D3 BioLegend 677807 AF647 AB_2616801 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
P08670 Primary Antibody Vimentin Human Mouse IgG2a O91D3 BioLegend 677807 AF647 AB_2616801 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 2
P08670 Primary Antibody Vimentin Human Mouse IgG2a O91D3 BioLegend 677807 AF647 AB_2616801 Stock Cell DIVE-IBEX FFPE Lung Cancer 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. Labels stromal cells in diverse FFPE tissues. Excellent signal and can be used at a 1:400-1:600 dilution.
  2. Labels stromal cells in diverse FFPE tissues. Excellent signal and can be used at a 1:400-1:600 dilution. Works beautifully on Cell DIVE imager with IBEX dye inactivation protocol.
  3. Labels stromal cells in lung FFPE tissue. Great signal and works very well on Cell DIVE imager with IBEX dye inactivation protocol.
Human liver: HLA-DR (cyan, catalog number 307620) and Vimentin (red, catalog number 677807)
Human jejunum: Hoechst (blue, catalog number 40046), CD54 (green, catalog number 353114), and Vimentin (magenta, catalog number 677807)
Human kidney: Hoechst (blue, catalog number 40046) and Vimentin (magenta, catalog number 677807)