‘Immunosequencing’ of heparan sulfate from human cell lines and rat kidney: the (GlcNS6S-IdoA2S) 3 motif, recognized by antibody NS4F5, is located towards.

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‘Immunosequencing’ of heparan sulfate from human cell lines and rat kidney: the (GlcNS6S-IdoA2S) 3 motif, recognized by antibody NS4F5, is located towards the non- reducing end by Xander M. R. van Wijk, Arie Oosterhof, Elly M. M. Versteeg, Els M. A. van de Westerlo, and Toin H. van Kuppevelt Biochem. J. Volume 461(3): August 1, 2014 ©2014 by Portland Press Ltd

Gradual reappearance of the amount of HS (and CS) after 4-deoxy-GlcNAc treatment MV3 cells were cultured for 3 days in the presence of 5 mM 4-deoxy-GlcNAc, after which the medium was replaced by medium without 4-deoxy-GlcNAc. Xander M. R. van Wijk et al. Biochem. J. 2014;461: ©2014 by Portland Press Ltd

Time-dependent reappearance of antibody-defined HS structures after 4-deoxy-GlcNAc treatment MV3 cells were cultured for 3 days in the presence of 5 mM 4-deoxy-GlcNAc, after which the medium was replaced by medium without 4-deoxy-GlcNAc. Xander M. R. van Wijk et al. Biochem. J. 2014;461: ©2014 by Portland Press Ltd

Peracetylated 4-deoxy-GlcNAc IC50 values for the inhibition of epitope-specific HS staining KOV3 cells were cultured for 3 days in the presence of 3–200 μM peracetylated 4-deoxy-GlcNAc, after which HS staining was evaluated by flow cytometry. Xander M. R. van Wijk et al. Biochem. J. 2014;461: ©2014 by Portland Press Ltd

Time-dependent removal of HS epitopes by heparinases Cultured MV3 melanoma cells were incubated with either 10 mIU/ml heparinase I or 20 mIU/ml heparinase III for 1–150 min. Xander M. R. van Wijk et al. Biochem. J. 2014;461: ©2014 by Portland Press Ltd

Concentration-dependent removal of HS epitopes by heparinases Cultured SKOV3 cells were incubated with different concentrations of either heparinase I or heparinase III for 5 h, and the loss of epitopes was assessed by flow cytometry. Xander M. R. van Wijk et al. Biochem. J. 2014;461: ©2014 by Portland Press Ltd

Time-dependent removal of HS epitopes from cortical kidney sections by heparinase III Kidney sections were incubated with heparinase III for 30 s to 10 min, and the loss of epitopes was assessed by immunohistochemistry. Xander M. R. van Wijk et al. Biochem. J. 2014;461: ©2014 by Portland Press Ltd

Time-dependent removal of HS epitopes from medullary kidney sections by heparinase III Kidney sections were incubated with heparinase III for 30 s to 2 h, and loss of epitopes was assessed by immunohistochemistry. Xander M. R. van Wijk et al. Biochem. J. 2014;461: ©2014 by Portland Press Ltd

Localized digestion of HS epitopes from medullary kidney sections by heparinase III Kidney sections were pre-loaded with digestion buffer, after which 5 μl of heparinase III was applied to one side of the section, resulting in diffusion of the enzyme and a... Xander M. R. van Wijk et al. Biochem. J. 2014;461: ©2014 by Portland Press Ltd