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
P20023 Primary Antibody CD21 Human Mouse IgG1 Bu32 BioLegend 354902 (Unconjugated) AF532 AB_2892739 Custom 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 1
P20023 Primary Antibody CD21 Human Mouse IgG1 Bu32 BioLegend 354902 (Unconjugated) AF532 AB_2892739 Custom IBEX2D Automated 1:4 Cytofix/Cytoperm Fixed Frozen Spleen 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 1
P20023 Primary Antibody CD21 Human Mouse IgG1 CR2/1952 Novus Biologicals NBP2-60733AF532 AF532 NA Stock Multiplexed 2D Imaging FFPE Tonsil NA 0.3% Triton-X-100 pH 6 for 40 minutes at 95C (AR6 Akoya Biosciences AR600250ML) NA No 0000-0003-4379-8967 NA 0000-0003-4379-8967 2
P20023 Primary Antibody CD21 Human Mouse IgG1 Bu32 BioLegend 354902 (Unconjugated) AF532 AB_2892739 Custom 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 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., “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. Custom conjugation by BioLegend
  2. Evaluated in human tonsil FFPE samples with CD20 (clone L26) to label B cell follicles. Does not label follicular dendritic cells in the lymph node. Recommend Abcam ab240987 (clone SP186) in place of this antibody.
Human lymph node: CD21 (magenta, based on catalog number 354902) and CD44 (cyan, based on catalog number 103002)
Human spleen: CD21 (red, based on catalog number 354902) and CD54 (blue, catalog number 353114)
Human spleen: Ki-67 (yellow, catalog number 561277) and CD21 (cyan, based on catalog number 354902)