| 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 |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| P02462 | Primary Antibody | Collagen IV | Human | Rabbit | IgG | Polyclonal | Abcam | ab6586 | Unconjugated | AB_305584 | Stock | IBEX2D Manual | 1:4 Cytofix/Cytoperm Fixed Frozen | Lymph Node | NA | 0.3% Triton-X-100 | NA | NA | Yes | 0000-0003-4379-8967 [2, 1] | NA | 0000-0003-4379-8967 | |
| P02463 | Primary Antibody | Collagen IV | Mouse | Rabbit | IgG | Polyclonal | Abcam | ab19808 | Unconjugated | AB_445160 | Stock | IBEX2D Manual | 1:4 Cytofix/Cytoperm Fixed Frozen | Spleen | NA | 0.3% Triton-X-100 | NA | NA | Yes | 0000-0003-4379-8967 [2] | NA | 0000-0003-4379-8967 | 1 |
| P02463 | Primary Antibody | Collagen IV | Mouse | Rabbit | IgG | Polyclonal | Abcam | ab19808 | Unconjugated | AB_445160 | Stock | IBEX2D Manual | 1:4 Cytofix/Cytoperm Fixed Frozen | Thymus | NA | 0.3% Triton-X-100 | NA | NA | Yes | 0000-0003-4379-8967 [2] | NA | 0000-0003-4379-8967 | |
| P02463 | Primary Antibody | Collagen IV | Mouse | Rabbit | IgG | Polyclonal | Abcam | ab19808 | Unconjugated | AB_445160 | Stock | IBEX2D Manual | 1:4 Cytofix/Cytoperm Fixed Frozen | Lung | NA | 0.3% Triton-X-100 | NA | NA | Yes | 0000-0003-4379-8967 [2] | NA | 0000-0003-4379-8967 | 1 |
| P02463 | Primary Antibody | Collagen IV | Mouse | Rabbit | IgG | Polyclonal | Abcam | ab19808 | Unconjugated | AB_445160 | Stock | IBEX2D Manual | 1:4 Cytofix/Cytoperm Fixed Frozen | Lymph Node | NA | 0.3% Triton-X-100 | NA | NA | Yes | 0000-0003-4379-8967 [2]; 0000-0003-1118-7432 | NA | 0000-0003-4379-8967 | |
| P02462 | Primary Antibody | Collagen IV | Human | Rabbit | IgG | Polyclonal | Abcam | ab6586 | Unconjugated | AB_305584 | Stock | IBEX2D Automated | 1:4 Cytofix/Cytoperm Fixed Frozen | Lymph Node | 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 | |
| P02462 | Primary Antibody | Collagen IV | Human | Rabbit | IgG | Polyclonal | Abcam | ab6586 | Unconjugated | AB_305584 | 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
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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)
| Human lymph node: Hoechst (blue, catalog number 40046) and Collagen IV (red, catalog number ab6586) |
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- Unconjugated primary antibody detected with goat anti-rabbit IgG AF700 (A-21038).
| Human jejunum: CD3 (red, catalog number 300448), Collagen IV (white, catalog number ab6586 and A-21038), and EpCAM (blue, catalog number 324228) |
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