LNs were collected and embedded in OCT Compound (Tissue-Tek). Cryostat sections (6 um thick) were mounted on Poly-lysine microscope slides (Thermo Scientific). Brightfield and DiD images of cryostat sections were acquired using Nikon Eclipse Ti-5 before acetone fixation. Sections were subsequently dried and fixed in cold acetone for 5 min at −20° C, blocked with 3% BSA in PBS blocking buffer at room temperature for 30 min, and incubated with anti-CD169 (ThermoScientific) and SIGNR1 (eBioSciences) antibodies diluted in the blocking buffer for 1 hour. Primary antibodies were detected with Goat-anti-Rat-AF488 (against CD169, BioLegend) and Goat-Anti-Hamster IgG-568 (against SIGNR1, Abcam). Images were acquired with the Nikon Eclipse Ti-5 microscope, using 4× objective lens and NIS-Elements D3.2 Software and processed with Fiji software.
Signr1
Manufactured by Thermo Fisher Scientific
SIGNR1 is a lab equipment product manufactured by Thermo Fisher Scientific. It functions as a carbohydrate-binding receptor that recognizes specific sugar molecules.
Automatically generated - may contain errors
2 protocols using signr1
1
Immunohistochemical Staining of Mouse Lymph Nodes
2
Fluorescent Labeling of Proteins and Antibodies
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Recombinant gp120 and HEL were conjugated to fluorescent (Alexa Fluor 488, 594, or
647) with the Microscale Protein Labeling Kit (Molecular probe, Thermo Fisher
Scientific). Antibodies against CD169 (3D6.112, BioLegend, San Diego, CA), SIGN-R1
(eBio22D1, eBiosciences, San Diego, CA), LYVE-1 (Clone# 223322, R&D
System, Minneapolis, MN) and ER-TR7 (ER-TR7, AbD serotec, Bio-Rad Laboratories,
Hercules, CA) were conjugated to Alexa Fluor 488, 594, or 647 with the Antibody
Labeling Kits (Molecular probe). Labeling reactions, conjugates purification, and
determination of degree of labeling were performed following the company manuals.
6–10-week-old recipient, anesthetized mice were injected with fluorescent
labeled materials for intravital imaging or section imaging by tail base
injection.
647) with the Microscale Protein Labeling Kit (Molecular probe, Thermo Fisher
Scientific). Antibodies against CD169 (3D6.112, BioLegend, San Diego, CA), SIGN-R1
(eBio22D1, eBiosciences, San Diego, CA), LYVE-1 (Clone# 223322, R&D
System, Minneapolis, MN) and ER-TR7 (ER-TR7, AbD serotec, Bio-Rad Laboratories,
Hercules, CA) were conjugated to Alexa Fluor 488, 594, or 647 with the Antibody
Labeling Kits (Molecular probe). Labeling reactions, conjugates purification, and
determination of degree of labeling were performed following the company manuals.
6–10-week-old recipient, anesthetized mice were injected with fluorescent
labeled materials for intravital imaging or section imaging by tail base
injection.
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