Cells cultured on permeable inserts were treated with 15 mU/mL of Heparinase III (New England Biolabs) or 2.5 mU/mL Chondroitinase ABC (R&D Systems) for 1 h at 37 °C [13 (link)]. Cells were then stimulated with HBP and TEER, and HRP passage was measured after 1.5 h.
Heparinase 3
Manufactured by New England Biolabs
Sourced in United Kingdom
Heparinase III is an enzyme used for the cleavage of heparin and related glycosaminoglycans. It functions by catalyzing the eliminative cleavage of heparin and heparan sulfate.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using heparinase 3
1
Enzymatic Barrier Modulation in Cell Cultures
2
Proteoglycan Isolation and Enzymatic Characterization
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Proteoglycans were isolated from the CM by precipitation with cationic detergent cetylpyridinium chloride, and the precipitates were dissolved in isopropanol. The polyanionic macromolecules were precipitated by adding sodium acetate-saturated ethanol and washed in ethanol. The extract was dissolved in ammonium acetate and then treated with chondroitinase AC (ChAC, Sigma-Aldrich), chondroitinase B (ChB, Sigma-Aldrich), chondroitinase ABC (ChABC, Sigma-Aldrich), heparinase I (NEB, Ipswich, MA, UK), heparinase II (NEB), or heparinase III (NEB) for further analysis.
3
Heparinase Treatment for GPC4 Quantification
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Samples containing Nluc-GPC4 were treated with Heparinase II (NEB P0735S) and Heparinase III (NEB P0737S) overnight at 37°C, then subject to DEAE pulldown described above, before luciferase activity was assayed. Untreated samples were similarly incubated at 37°C and used as controls for Heparinase treatment for the DEAE pulldown and luciferase assay.
4
Krebs-Henseleit Buffer Perfusion Protocol
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Krebs–Henseleit buffer, Nω-nitro-l -arginine methyl ester (L-NAME), apocynin were purchased from Sigma-Aldrich (St Louis, MO). Bovine serum albumin (BSA) was purchased from Proliant (Ankeny, IA). BSA-FITC conjugate was purchased from ThermoFisher Scientific (Waltham, MA). Snapwells were purchased from Corning Life Sciences (Corning, NY). Albumin Blue Fluorescent kit was purchased from Active Motif (Carlsbad, CA). Heparinase III was purchased from New England Biolabs (Ipswich, MA).
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