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 Sheep IgG (H+L) Human Donkey IgG Polyclonal Thermo Fisher Scientific A-21102 AF680 AB_2535755 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 [2, 1] NA 0000-0003-4379-8967  
NA Secondary Antibody Sheep IgG (H+L) Human Donkey IgG Polyclonal Thermo Fisher Scientific A-21102 AF680 AB_2535755 Stock IBEX2D Manual 1:4 Cytofix/Cytoperm Fixed Frozen Spleen NA 0.3% Triton-X-100 NA 1 mg/ml LiBH4 15 minutes Yes 0000-0003-4379-8967 [2, 1] NA 0000-0003-4379-8967  
NA Secondary Antibody Sheep IgG (H+L) Human Donkey IgG Polyclonal Thermo Fisher Scientific A-21102 AF680 AB_2535755 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 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., “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. This antibody can be used to evaluate unconjugated sheep primary antibodies in a wide range of tissues. Be careful when using unconjugated primary antibodies as IBEX protocol does not strip antibodies. Place antibodies in first cycle and use direct conjugates in later cycles. Careful when combining with goat, rabbit, and other unconjugated primary antibodies as secondary antibodies may not be cross-adsorbed.
Human spleen: CD49a (cyan, catalog number 328310) and Fibrinogen (red, catalog number ab118533 and A-21102)