20 mg/ml solutions of Copaxone® (glatiramer acetate) 1 ml pre-filled syringes from Teva Neuroscience, Inc. (Kansas City, MO) were donated by the University of Kansas Medical Center. For peptide synthesis, Fmoc-L-phenylalanine 4-alkoxybenzyl alcohol resin (0.3–0.8 meq/g, 100–200 mesh) and all amino acids were purchased from Chem-Impex International Inc. (Wood Dale, IL). Fmoc-NH-(PEG)4-CH2COOH (5,8,11,14-Tetraoxa-2-azahexadecanedioic acid) was purchased from PurePEG, LLC (San Diego, CA). Myelin proteolipid protein (PLP) epitope (HSLGKWLGHPDKF) and 9-unit poly-lysine were synthesized and purchased from Biomatik USA, LLC (Wilmington, DE). Scissor Cartridge packs containing HA based extracellular matrix (ECM) were obtained from Pion Inc. (Billerica, MA). Sulfo-Cyanine7 NHS ester was purchased from Lumiprobe (Hunt Valley, MD). PEG amine (5kDa) was obtained from Creative PEGworks (Chapel Hill, NC). All other analytical grade chemicals and reagents were purchased from MilliporeSigma (St. Louis, MO) and Fisher Scientific (Pittsburgh, PA) and were used as received. All mice used for the study were maintained in sterile housing under the veterinary supervision of the University of Kansas Animal Care Unit. All procedures were approved by the Institutional Animal Care and Use Committee.
Sulfo cyanine7 nhs ester
Manufactured by Lumiprobe
Sulfo-Cyanine7 NHS ester is a water-soluble cyanine dye with an N-hydroxysuccinimide (NHS) ester reactive group. The Sulfo-Cyanine7 fluorophore has an absorption maximum at 743 nm and an emission maximum at 767 nm, making it suitable for near-infrared (NIR) imaging applications.
Automatically generated - may contain errors
Lab products found in correlation
Polyacrylamide gel, by Bio-Rad (2 mentions)
Recombinant human il 15, by BioLegend (2 mentions)
Dbco nhs ester, by Vector Laboratories (2 mentions)
Recombinant mouse scil 12, by Miltenyi Biotec (2 mentions)
Recombinant human il 2, by Thermo Fisher Scientific (2 mentions)
Ifnγ elisa, by R&D Systems (1 mentions)
Ifn γ elisa, by BioLegend (1 mentions)
Odyssey clx, by LI COR (1 mentions)
4 protocols using sulfo cyanine7 nhs ester
1
Glatiramer Acetate Formulation and Synthesis
2
Cytokine-Conjugated Polymer Nanomedicines
3
Cytokine-mediated T cell activation
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Unless otherwise specified,
reagents were used as received without further purification. Recombinant
human IL-2 (200-02, Peprotech), recombinant human IL-15 (570308, Biolegend),
recombinant mouse scIL-12 (130-096, Miltenyi Biotec), and recombinant
human IL-12 (CT050-HNAH, Sino Biological). Sulfo-Cyanine7 NHS ester
(Lumiprobe), DBCO-NHS ester (1160, Click Chemistry Tools), NHS-PEG-TCO
ester (A137-10, Click Chemistry Tools), poly(ethylene glycol)methyl
ether DBCO (20 kDa, A120-100, Click Chemistry Tools), and poly(ethylene
glycol)methyl ether azide (20 kDa, Nanocs). Polyacrylamide gels (Bio-Rad,
4–16 wt %). IFNγ ELISA (430104, Biolegend). CD19 antibody
(BD Biosciences Clone SJ25C1) and PD-L1 antibody (eBioscience Clone
MIH1).
reagents were used as received without further purification. Recombinant
human IL-2 (200-02, Peprotech), recombinant human IL-15 (570308, Biolegend),
recombinant mouse scIL-12 (130-096, Miltenyi Biotec), and recombinant
human IL-12 (CT050-HNAH, Sino Biological). Sulfo-Cyanine7 NHS ester
(Lumiprobe), DBCO-NHS ester (1160, Click Chemistry Tools), NHS-PEG-TCO
ester (A137-10, Click Chemistry Tools), poly(ethylene glycol)methyl
ether DBCO (20 kDa, A120-100, Click Chemistry Tools), and poly(ethylene
glycol)methyl ether azide (20 kDa, Nanocs). Polyacrylamide gels (Bio-Rad,
4–16 wt %). IFNγ ELISA (430104, Biolegend). CD19 antibody
(BD Biosciences Clone SJ25C1) and PD-L1 antibody (eBioscience Clone
MIH1).
4
In Vivo Biodistribution of Fluorescent DNA Conjugates
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Near-infrared-dye-labelled agents were used to minimize interference from autofluorescent background in vivo. BALB/c mice were intravenously injected with Cy5-labelled modified or native DNA molecules (1 nmol) and urine samples were collected at 30 min and 1, 2, 3, 4 h after injection. Nanobody–DNA conjugates were coupled with sulfo-Cyanine7 NHS ester (Lumiprobe, 2 dye equiv. of protein), reacted overnight, purified by spin filtration and injected intravenously into PC-3 tumour-bearing nude mice. After 24 h, mice were euthanized and necropsy was performed to remove the tumours, lungs, heart, kidneys, liver and spleen. Urine, blood and organs were scanned using IVIS and Odyssey CLx imaging systems (LI-COR). Organ fluorescence was quantified by ImageStudio in the Odyssey CLx. Blood circulatory kinetics were monitored in BALB/c mice by serial blood draws at 10 min, 30 min, 2 h and 3 h after intravenous injection of Cy5-labelled DNA or PEG at 1 nmol dye per mouse. Blood for pharmacokinetics measurements was collected using tail vain bleeds. Blood was diluted in PBS with 5 mM EDTA to prevent clotting, centrifuged for 5 min at 5,000g, and fluorescent reporter concentration was quantified in 384-well plates relative to standards on the Odyssey CLx.
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