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
Q7ZVF2 Primary Antibody Glutamine synthetase Zebrafish Mouse IgG1 1B6G4 Proteintech CL488-66323 CoraLite Plus AF488 AB_2883309 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 1 mg/ml LiBH4 30 minutes + light Yes 0009-0000-2047-4228 [1] NA 0009-0000-2047-4228 1
A0A8C6NQP0 Primary Antibody Glutamine synthetase Turquoise Killifish Mouse IgG1 1B6G4 Proteintech CL488-66323 CoraLite Plus AF488 AB_2883309 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 1 mg/ml LiBH4 30 minutes + light Yes 0009-0000-2047-4228 [1] NA 0009-0000-2047-4228  
P51121 Primary Antibody Glutamine synthetase African Clawed Frog (Xenopus laevis) Mouse IgG1 1B6G4 Proteintech CL488-66323 CoraLite Plus AF488 AB_2883309 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 1 mg/ml LiBH4 30 minutes + light 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. Longer dye inactivation conditions reflect requirements to inactivate antibodies conjugated with FlexAble kits imaged in the same cycle with this GFAP AF488 antibody. See methods section of bioRxiv. CoraLite Plus AF488 conjugates dye inactivate within 15 minutes of 1 mg/ml LiBH4 (see fluorescent probes).