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
P01731 Primary Antibody CD8 Mouse Rat IgG2a 53-6.7 BioLegend 100724 AF647 AB_389326 Stock IBEX2D Manual 1:4 Cytofix/Cytoperm Fixed Frozen Lung NA 0.3% Triton-X-100 NA 1 mg/ml LiBH4 15 minutes Yes 0000-0003-4379-8967 [1] NA 0000-0003-4379-8967 1
P01732 Primary Antibody CD8 Human Mouse IgG1 C8/144B BioLegend 372906 AF647 AB_2650712 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, 0009-0006-9784-2694 NA 0000-0003-4379-8967 2
P01732 Primary Antibody CD8 Human Mouse IgG1 C8/144B BioLegend 372906 AF647 AB_2650712 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
P01732 Primary Antibody CD8 Human Mouse IgG1 C8/144B BioLegend 372906 AF647 AB_2650712 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 4
P01732 Primary Antibody CD8 Human Mouse IgG1 C8/144B BioLegend 372906 AF647 AB_2650712 Stock Cell DIVE-IBEX FFPE Lymph Node Healthy 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 [2, 3] NA 0000-0003-4379-8967 5

Publications

  1. 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

  2. A. J. Radtke et al., “Multi-omic profiling of follicular lymphoma reveals changes in tissue architecture and enhanced stromal remodeling in high-risk patients”, Cancer Cell, 42(3):444-463, 2024, doi: 10.1016/j.ccell.2024.02.001.

  3. A. J. Radtke, “OMAP-18: Organ Mapping Antibody Panel (OMAP) for Multiplexed Antibody-Based Imaging of Human Lymph Node with Cell DIVE-IBEX”, Human Reference Atlas, 2024, doi: 10.48539/HBM356.HGHM.686.

Additional Notes

  1. Validated by Evelyn Kandov (No ORCID)
  2. Evaluated in human tonsil FFPE samples with CD20 (clone L26) and Hoechst (nuclear marker). Antibody labels T cells. Preferred clone and conjugate for FFPE samples.
  3. Antibody labels T cells. Preferred clone and conjugate for FFPE samples. Works beautifully on Cell DIVE imager with IBEX dye inactivation protocol.
  4. Antibody labels cytotoxic T cells in various human FFPE tissues. Great signal in the lung and works well with the Cell DIVE imager and IBEX dye inactivation protocol.
  5. CD8 AF647 (BioLegend 372906) was used at a 1:50 dilution. Works great!
Human normal LN: CD3 (blue, catalog number ab208514) and CD8 (cyan, catalog number 372906)
Human normal LN: CD3 (blue, catalog number ab208514), CD8 (cyan, catalog number 372906), and CD4 (red, catalog number ab196372)