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
NA Secondary Antibody Rabbit IgG (H+L) Human Goat IgG Polyclonal Thermo Fisher Scientific A-21038 AF700 AB_2535709 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  
NA Secondary Antibody Rabbit IgG (H+L) Mouse Goat IgG Polyclonal Thermo Fisher Scientific A-21038 AF700 AB_2535709 Stock IBEX2D Manual 1:4 Cytofix/Cytoperm Fixed Frozen Thymus NA 0.3% Triton-X-100 NA 1 mg/ml LiBH4 15 minutes Yes 0000-0003-4379-8967 [3] NA 0000-0003-4379-8967  
NA Secondary Antibody Rabbit IgG (H+L) Mouse Goat IgG Polyclonal Thermo Fisher Scientific A-21038 AF700 AB_2535709 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 [3] NA 0000-0003-4379-8967 1
NA Secondary Antibody Rabbit IgG (H+L) Mouse Goat IgG Polyclonal Thermo Fisher Scientific A-21038 AF700 AB_2535709 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] NA 0000-0003-4379-8967  
NA Secondary Antibody Rabbit IgG (H+L) Human Goat IgG Polyclonal Thermo Fisher Scientific A-21038 AF700 AB_2535709 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  
NA Secondary Antibody Rabbit IgG (H+L) Human Goat IgG Polyclonal Thermo Fisher Scientific A-21038 AF700 AB_2535709 Stock Multiplexed 2D Imaging 1:4 Cytofix/Cytoperm Fixed Frozen Jejunum NA 0.3% Triton-X-100 NA NA Yes 0000-0003-4379-8967 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., “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. Validated by Evelyn Kandov (No ORCID)
  2. Used to detect unconjugated primary antibody (Abcam, ab6586). | Human jejunum: CD3 (red, catalog number 300448), Collagen IV (white, catalog number ab6586 and A-21038), and EpCAM (blue, catalog number 324228) | |:——-:| | |