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
NA	Primary Antibody	HuC/D	Zebrafish	Mouse	IgG2b	16A11	Thermo Fisher Scientific	A21271	Unconjugated	AB_221448	Stock	IBEX2D Manual	4% PFA Fixed Frozen	Retina	GFP transgenic line	0.1% Triton-X-100	pH 6 (10 mM Sodium Citrate) for 20 minutes in a pressure cooker	NA	Yes	0009-0000-2047-4228 [1]	NA	0009-0000-2047-4228	1
NA	Primary Antibody	HuC/D	Turquoise Killifish	Mouse	IgG2b	16A11	Thermo Fisher Scientific	A21271	Unconjugated	AB_221448	Stock	IBEX2D Manual	4% PFA Fixed Frozen	Retina	NA	0.1% Triton-X-100	pH 6 (10 mM Sodium Citrate) for 20 minutes in a pressure cooker	NA	Yes	0009-0000-2047-4228 [1]	NA	0009-0000-2047-4228
NA	Primary Antibody	HuC/D	African Clawed Frog (Xenopus laevis)	Mouse	IgG2b	16A11	Thermo Fisher Scientific	A21271	Unconjugated	AB_221448	Stock	IBEX2D Manual	4% PFA Fixed Frozen	Retina	NA	0.1% Triton-X-100	pH 6 (10 mM Sodium Citrate) for 20 minutes in a pressure cooker	NA	Yes	0009-0000-2047-4228 [1]	NA	0009-0000-2047-4228

Publications

1. A. Kothurkar et al., “Iterative Bleaching Extends Multiplexity (IBEX) imaging facilitates simultaneous identification of all cell types in the vertebrate retina”, bioRxiv, 2024, doi: 10.1101/2024.02.28.582563.

Additional Notes

1. Prior to IBEX imaging, antigen retrieval was performed by heating the slides for 20 minutes in 10 mM sodium citrate (pH 6). This step quenched the signal in most of the zebrafish GFP transgenic lines tested, but the GFP can be boosted in later rounds of immunolabelling using a primary antibody against GFP.