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
P01730 Primary Antibody CD4 Human Mouse IgG1 RPA-T4 BioLegend 300519 AF488 AB_389311 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  
P06332 Primary Antibody CD4 Mouse Rat IgG2a RM4-5 BD Biosciences 557667 AF488 AB_396779 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] NA 0000-0003-4379-8967  
P01730 Primary Antibody CD4 Human Rabbit IgG EPR6855 Abcam ab196372 AF488 AB_2889191 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 NA 0000-0003-4379-8967 1
P01730 Primary Antibody CD4 Human Rabbit IgG EPR6855 Abcam ab196372 AF488 AB_2889191 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 2
P01730 Primary Antibody CD4 Human Rabbit IgG EPR6855 Abcam ab196372 AF488 AB_2889191 Stock Cell DIVE-IBEX FFPE Lymph Node Follicular Lymphoma 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 [3] NA 0000-0003-4379-8967  
P01730 Primary Antibody CD4 Human Rabbit IgG EPR6855 Abcam ab196372 AF488 AB_2889191 Stock Cell DIVE-IBEX FFPE Lymph Node 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 [3] NA 0000-0003-4379-8967  

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

  3. 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.

Additional Notes

  1. Antibody labels T helper cells and macrophages if used at a high concentration (1:25). Dual antigen retrieval (pH 6 and 9) is required for this antibody.
  2. Antibody labels T helper cells and macrophages if used at a high concentration (1:25). Dual antigen retrieval (pH 6 and 9) is required for this antibody. Works beautifully on Cell DIVE imager with IBEX dye inactivation protocol.
Human lymph node: CD163 (red, catalog number 333620) and CD4 (blue, catalog number 300519)
Human normal LN: BCL2 (cyan, catalog number ab236221 and A-31573), CD3 (magenta, catalog number ab208514), CD4 (blue, catalog number ab196372), CD21 (yellow, catalog number ab306325)
Human FL LN: BCL2 (cyan, catalog number ab236221 and A-31573), CD3 (magenta, catalog number ab208514), CD4 (blue, catalog number ab196372), CD21 (yellow, catalog number ab306325)