Nanoparticles were incubated with neuraminidase A (Neu A, New England biolabs, Inc., Beverly, MA, USA), neuraminidase S (Neu S, New England biolabs, Inc.), or PNGase F (New England biolabs, Inc.) at 37 °C for 18 h. Proteins were transferred to a polyvinylidene fluoride membrane (Thermo Fisher Scientific, Waltham, MA, USA). After blocking with 5% (w/v) bovine serum albumin in Tris-buffered saline with 0.05% TWEEN 20 (TBST) for 1 h at 25 °C, membranes were probed with monoclonal mouse anti-flag (1:1000, Sigma-Aldrich, St. Louis, MO, USA) antibodies, biotinylated Sambucus nigra agglutinin (SNA, Vector Laboratories, Burlingame, CA, USA), or biotinylated Maackia amurensis lectin-I (MAL-I, Vector Laboratories, Burlingame, CA, USA) as a primary staining, and then with horseradish peroxidase-conjugated goat anti-mouse IgG (Bioss Antibodies, Woburn, MA, USA) and streptavidin at 1:5000 dilution in TBST. Membranes were developed using ECL substrate (Thermo Fisher Scientific, Waltham, MA, USA) and signals were detected with Fusion Solo X (Vilber, Paris, France).
Neu s
Manufactured by New England Biolabs
Sourced in United States
Neu S is a laboratory equipment product that functions as a DNA restriction endonuclease enzyme. It recognizes and cleaves specific DNA sequences, facilitating DNA manipulation and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Pngase f, by New England Biolabs (2 mentions)
Ecl substrate, by Thermo Fisher Scientific (1 mentions)
Polyvinylidene fluoride membrane, by Thermo Fisher Scientific (1 mentions)
Mouse monoclonal anti flag, by Merck Group (1 mentions)
Horseradish peroxidase conjugated goat anti mouse igg, by Bioss Antibodies (1 mentions)
Neu a, by New England Biolabs (1 mentions)
Biotinylated sambucus nigra agglutinin sna, by Vector Laboratories (1 mentions)
Fusion solo x, by Vilber (1 mentions)
Biotinylated maackia amurensis lectin 1 mal 1, by Vector Laboratories (1 mentions)
2 protocols using neu s
1
Glycan Profiling of Nanoparticles
2
Flow Cytometry and Cell Viability Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
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For flow cytometry assays, cells were incubated with antibodies as per the manufacturer’s instructions. Cells treated with PNGaseF or NeuS (NEB) were incubated with 1 U of enzyme for 1 hour at 37°C before staining with antibodies. For cell viability assays, cells were seeded at a density of 5000 cells per well. Twenty-four hours later, cells were treated with a range of concentrations (2.5, 5, 10, 25, and 50 μM) of 5-FU and saline solution (control). Ninety-six hours after treatment, cells were assessed for viability using crystal violet. The absorbance measured at 600 nm was normalized to the saline solution control.
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