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 BioLegend 100234 BV510 AB_2562555 Stock IBEX2D Manual 1:4 Cytofix/Cytoperm Fixed Frozen Lymph Node NA 0.3% Triton-X-100 NA 1 mg/ml LiBH4 15 minutes + light Yes 0000-0003-4379-8967 [2] NA 0000-0003-4379-8967 1
P07766 Primary Antibody CD3 Human Mouse IgG1 UCHT1 BioLegend 300448 BV510 AB_2563468 Stock Multiplexed 2D Imaging 1:4 Cytofix/Cytoperm Fixed Frozen Lymph Node NA 0.3% Triton-X-100 NA NA Yes 0000-0003-4379-8967 [1] NA 0000-0003-4379-8967 2
P07766 Primary Antibody CD3 Human Mouse IgG1 UCHT1 BioLegend 300448 BV510 AB_2563468 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 3

Publications

  1. I. Sayin et al., “Spatial distribution and function of T follicular regulatory cells in human lymph nodes”, J Exp Med., 215(6):1531-1542, 2018, doi:10.1084/jem.20171940.

  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. Works but recommend other conjugates such as AF488, PE, or AF647
  2. Clone UCHT1 labels T cells in fixed frozen human tissues. Several conjugates are suitable for Multiplexed 2D Imaging; however, we recommend other conjugates (AF488, AF555, PE, iF594, AF647) for IBEX experiments. This conjugate can be used in human lymph nodes or tissues with low autofluorescence (see publication).
  3. Antibody labels T cells in jejunum but high autofluorescence observed with BV510 conjugates in this tissue. Background subtraction with channel arithmetics is recommended.
Human jejunum: CD3 (red, catalog number 300448), Collagen IV (white, catalog number ab6586 and A-21038), and EpCAM (blue, catalog number 324228)