| 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 |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| P22646 | Primary Antibody | CD3 | Mouse | Rat | IgG2b | 17A2 | BD Biosciences | 557869 | AF647 | AB_396912 | 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] | NA | 0000-0003-4379-8967 | 1 |
| P22646 | Primary Antibody | CD3 | Mouse | Rat | IgG2b | 17A2 | BD Biosciences | 557869 | AF647 | AB_396912 | Stock | IBEX2D Manual | 1:4 Cytofix/Cytoperm Fixed Frozen | Small Intestine | NA | 0.3% Triton-X-100 | NA | 1 mg/ml LiBH4 15 minutes | Yes | 0000-0003-4379-8967 [2] | NA | 0000-0003-4379-8967 | 1 |
| P07766 | Primary Antibody | CD3 | Human | Mouse | IgG1 | UCHT1 | BioLegend | 300416 | AF647 | AB_389332 | 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 [2, 1] | NA | 0000-0003-4379-8967 | 2 |
Publications
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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.
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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
- Validated by Evelyn Kandov (No ORCID)
- Clone UCHT1 labels T cells in fixed frozen human tissues. Several conjugates are suitable for Multiplexed 2D Imaging; however, we recommend the following conjugates for IBEX experiments: AF488, AF555, PE, iF594, and AF647.