Posttranslational modification of proteins by beta-linked N-acetylglucosamine (beta-GlcNAc) via the hydroxyl moieties on serine or threonine residues is termed O-linked beta-GlcNAc or simply O-GlcNAc. O-GlcNAc is one of the most abundant posttranslational modifications within the nucleocytoplasmic compartments of all animals and plants. Unlike other types of protein glycosylations, O-GlcNAc occurs exclusively within the nuclear and cytoplasmic compartments and is generally not further modified to form more elongated structures. In addition, O-GlcNAcylation is a highly dynamic and reversible process. The O-GlcNAc transferase (OGT) attaches O-GlcNAc to proteins at specific serine or threonine residues, while O-GlcNAcase catalyzes the removal/hydrolysis of O-GlcNAc from proteins. In fact, a dynamic interplay between O-GlcNAcylation and serine/threonine phosphorylation plays an important role in regulating cellular signaling. Tau and RNA polymerase II (Pol II) are two well known proteins that undergo modification by O-GlcNAcylation. In Alzheimer's diseased human brains, tau becomes extensively phosphorylated and less O-GlcNAcylated. Similarly, O-GlcNAc is removed and replaced with O-phosphate on the Poly II CTD when the elongation phase of transcription is initiated.
Synonyms: O-GlcNAc, beta-O-GlcNAc, O-Linked N-Acetylglucosamine, beta-O-linked N-acetylglucosamine
Application: Western Blotting Analysis: 1.0 µg/mL from a representative lot detected O-GlcNAcylated proteins in 7.5-15 µg of wild-type mouse embryonic fibroblast (MEF) lysate, but not O-GlcNAc transferase/OGT-deficient MEF lysate (Courtesy of Dr. Natasha Zachara and Gokben Yildirir, M.S.).ELISA Analysis: A representative lot detected RNA polymerase II C-terminal domain (CTD) peptide (YSPTSPS) with a single O-GlcNAcylated serine or threonine, but not the corresponding unmodified peptide (Comer, F.I., et al. (2001). Anal. Biochem. 293(2):169-177).Immunoprecipitation Analysis: A representative lot immunoprecipitated O-GlcNAcylated proteins from human pluripotent stem cells (hPSCs) (Maury, J.J., et al. (2013). Stem Cell Res. 11(2):926-937).Immunoprecipitation Analysis: A representative lot immunoprecipitated O-GlcNAcylated proteins from HeLa cell extracts (Comer, F.I., et al. (2001). Anal. Biochem. 293(2):169-177).Western Blotting Analysis: A representative lot detected similar level of cellular O-GlcNAcylation in undifferentiated, differentiating and terminally differentiated human pluripotent stem cells (hPSCs) (Maury, J.J., et al. (2013). Stem Cell Res. 11(2):926-937).Western Blotting Analysis: A representative lot detected BSA-conjugated RNA polymerase II C-terminal domain (CTD) peptide (YSPTSPS) with beta-O-linked GlcNAc, but not alpha-O-linked GlcNAc, nor the corresponding unmodified peptide. The presence of GlcNAc, but not GalNAc, abolished the detection of the target bands (Comer, F.I., et al. (2001). Anal. Biochem. 293(2):169-177).Western Blotting Analysis: A representative lot detected O-GlcNAcylated proteins in HeLa nuclear extract, as well as O-GlcNAcylated proteins purified from HeLa nuclear & cytosolic extract by wheat germ agglutinin (WGA) column. Antibody blocking by mmunogen peptide prior to immunoblotting abolished target bands detection (Comer, F.I., et al. (2001). Anal. Biochem. 293(2):169-177).Western Blotting Analysis: A representative lot detected an upregulation of O-GlcNAcylated proteins in Jurkat cells treated with the glucosaminidase inhibitor PUGNAc and the hexosamine pathway intermediate glucosamine (Comer, F.I., et al. (2001). Anal. Biochem. 293(2):169-177).
Other Notes: Concentration: Please refer to lot specific datasheet.