O-GlcNAc Antibody (HGAC85) - BSA Free

Western Blot: O-GlcNAc Antibody (HGAC85) [NB300-614] - alpha-DG-Fc was transiently transfected with or without AGO61 into COS1 cells. alpha-DG-Fc recombinant proteins were collected from cell lysates and culture media using protein A resin and analyzed for laminin overlay and Western blot using anti-Fc and anti-O-GlcNAc antibodies. Image collected and cropped by CiteAb from the following publication (nature.com/articles/srep03288), licensed under a CC-BY license.

Immunohistochemistry-Paraffin: O-GlcNAc Antibody (HGAC85) [NB300-614] - IHC staining of O-GlcNAc in mouse colon using DAB with hematoxylin counterstain.

Western Blot: O-GlcNAc Antibody (HGAC85) [NB300-614] - Western blot analysis of O-GlcNAc expression in 1) HeLa, 2) NTERA-2, 3) PC-12 and 4) COS-7 cell lysates using untreated antibody on the left side and antibody preincubated with 50 uM GlcNAc (see Notes).

Western Blot: O-GlcNAc Antibody (HGAC85) [NB300-614] - AGO61 modifies GlcNAc residues at specific sites on alpha -DG.(a) alpha -DG-Fc was transiently transfected with or without AGO61 into COS1 cells. alpha -DG-Fc recombinant proteins were collected from cell lysates & culture media using protein A resin & analyzed for laminin overlay & Western blot using anti-Fc & anti-O-GlcNAc antibodies. (b) alpha -DG373-HALO & its mutant T317A/T319A were transiently transfected with or without LARGE-myc into COS7 cells. HALO-fused proteins were collected from medium using HALO resin followed by digestion with TEV protease & then analyzed by laminin overlay. Cell lysates were analyzed for the expression of HALO-fused proteins & LARGE-myc by Western blot using anti-HALO & anti-myc antibodies. The full-length blots with anti-HALO & anti-myc antibodies are presented in Supplementary Figs. S7f & S7g, respectively. (c) alpha -DG373-HALO & its mutant T317A/T319A were transiently transfected with or without AOG61 into COS7 cells. HALO-fused proteins were collected from the cell lysates using HALO resin followed by digestion with TEV protease & then analyzed by Western blot using an anti-O-GlcNAc antibody (CTD110.6). The cell lysates were analyzed for the expression of HALO-fused proteins & AGO61 by Western blot using anti-HALO & anti-AGO61 antibodies. The full-length blots with anti-HALO & anti-AGO61 antibodies are presented in Supplementary Figs. S7h & S7i, respectively. Image collected & cropped by CiteAb from the following publication (//www.nature.com/articles/srep03288), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, All, Pm
• Applications: WB, ChIP, ELISA, ICC/IF, IHC, IP, ChIP
• Clone: HGAC85
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Format: BSA Free

O-GlcNAc Antibody (HGAC85) - BSA Free Summary

• Description: Novus Biologicals Mouse O-GlcNAc Antibody (HGAC85) - BSA Free (NB300-614) is a monoclonal antibody validated for use in IHC, WB, ELISA, ICC/IF, IP and ChIP. Anti-O-GlcNAc Antibody: Cited in 11 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: Heat-killed, pepsin-treated group A streptococci (Streptococcus pyogenes)
• Specificity: Recognizes beta-1,3 linked O-linked N-acetylglucosamine (O-GlcNAc) residues of streptococcal group A carbohydrate as well as O-GlcNAc glycosylated proteins
• Isotype: IgG3 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Purity: Unpurified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation (ChIP) 1:10-1:500
  • Chromatin Immunoprecipitation 1:10-1:500. Use reported in scientific literature (PMID 20368426)
  • ELISA 1:100-1:2000
  • Immunocytochemistry/ Immunofluorescence 1:100 - 1:200. Use reported in scientific literature (Hamiel CR et al)
  • Immunohistochemistry 1:200. Use reported in scientific literature (Zhang , et al)
  • Immunohistochemistry-Paraffin 1:200
  • Immunoprecipitation 1:10-1:500. Use reported in scientific literature (Hamiel CR et al)
  • Western Blot 1:500-1:1000
• Application Notes: In Western blot, this antibody detects several proteins representing O-GlcNAc glycoproteins. Immunofluorescence staining of O-GlcNAc in cells results in labeling of the nuclear envelope and pores, nucleolus, and cytoplasm. This staining pattern is consistent with other methods of detecting O-GlcNAc moieties. DO NOT USE WITH DILUENTS CONTAINING GLYCOSYLATED PROTEINS.

Reactivity Notes

All Species. Human reactivity reported in scientific literature (PMID: 28973823). Chlorocebus sabaeus (Green monkey) reported in PMID 24256719.

Packaging, Storage & Formulations

• Storage: Store at -20C. Avoid freeze-thaw cycles.
• Buffer: Ascites
• Preservative: 0.1% Sodium Azide
• Purity: Unpurified

Alternate Names for O-GlcNAc Antibody (HGAC85) - BSA Free

• GlcNAc
• O-linked N-acetylglucosamine

Background

O-GlcNAc (O-linked N-acetylglucosamine) is one of the most abundant posttranslational modifications on nuclear and cytoplasmic proteins. Many cellular proteins, including nuclear pore, oncogene, cytoskeletal, heat shock, viral and transcription regulatory proteins contain single O-GlcNAc residues attached to serine or threonine residues. This modification occurs via the O-GlcNAc transferase (OGT). There appears to be a competitive modification at these residues. Either the residues are glycosylated or they are phosphorylated. Because of the significance of phosphorylation in cancer research, such competition can be of major interest. O-GlcNAc bearing proteins tend to be found in multimeric complexes. This has led to the suggestion that O-GlcNAc glycosylation may also obscure phosphorylation sites and acts as a signaling mechanism or mediator of signaling. Recent research has revealed that the O-GlcNAcylation of the EGF repeats of Notch are modified via a novel O-GlcNAc transferase, EOGT1 (PMID: 22310717).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for O-GlcNAc Antibody (NB300-614)(11)

Publications using NB300-614

• Akinbiyi EO, Abramowitz LK, Bauer BL et al. Blocked O-GlcNAc cycling alters mitochondrial morphology, function, and mass Scientific Reports 2021-11-11 [PMID: 34764359] (Immunocytochemistry/ Immunofluorescence, Rat)
• Ishimura E, Nakagawa T, Moriwaki K et al. Augmented O-GlcNAcylation of AMP-activated kinase promotes the proliferation of LoVo cells, a colon cancer cell line. Cancer Sci. 2017-10-03 [PMID: 28973823] (WB, Human)
• Yagi H, Nakagawa N, Saito T et al. AGO61-dependent GlcNAc modification primes the formation of functional glycans on alpha-dystroglycan. Sci Rep. 2013-11-21 [PMID: 24256719] (WB, Primate)
• Love DC, Ghosh S, Mondoux MA et al. Dynamic O-GlcNAc cycling at promoters of Caenorhabditis elegans genes regulating longevity, stress, and immunity. Proc Natl Acad Sci U S A. 2010-04-01 [PMID: 20368426]
• Hamiel CR, Pinto S, Hau A et al. Glutamine enhances heat shock protein 70 expression via increased hexosamine biosynthetic pathway activity. Am J Physiol Cell Physiol. 2009-12-01 [PMID: 19776393]
• Akimoto Y, Kreppel LK, Hirano H, Hart GW. Localization of the O-linked N-acetylglucosamine transferase in rat pancreas. Diabetes. 1999-12-01 [PMID: 10580430] (ICC/IF, Rat)
• Zhang X, Bennett V. Identification of O-linked N-acetylglucosamine modification of ankyrinG isoforms targeted to nodes of Ranvier. J Biol Chem. 1996-12-01 [PMID: 8940148]
• Greenspan NS, Dacek DA, Cooper LJ. Fc region-dependence of IgG3 anti-streptococcal group A carbohydrate antibody functional affinity. I. The effect of temperature. J Immunol. 1988-12-15 [PMID: 3058803]
• Greenspan NS, Dacek DA, Cooper LJ. Cooperative binding of two antibodies to independent antigens by an Fc-dependent mechanism. FASEB J. 1989-08-01 [PMID: 2666233]
• Greenspan NS, Monafo WJ, Davie JM. Interaction of IgG3 anti-streptococcal group A carbohydrate (GAC) antibody with streptococcal group A vaccine: enhancing and inhibiting effects of anti-GAC, anti-isotypic, and anti-idiotypic antibodies. J Immunol. 1987-01-01 [PMID: 2431057]
• Turner JR, Tartakoff AM, Greenspan NS. Cytologic assessment of nuclear and cytoplasmic O-linked N-acetylglucosamine distribution by using anti-streptococcal monoclonal antibodies. Proc Natl Acad Sci USA. 1990-08-01 [PMID: 2116002] (ICC/IF, Rat)

FAQs for O-GlcNAc Antibody (NB300-614). (Showing 1 - 1 of 1 FAQs).

1. Is it possible to get your mAb without BSA or sodium azide? I am interested in NB300-614 and NB300-524 .
   • These antibodies are supplied in Sodium Azide, but if you are interested, we do provide kits to clean up the antibodies. Here is the link to our AbSelect Antibody Purification Kits .