Anti-O-Linked N-Acetylglucosamine antibody [HGAC85] - ChIP Grade (ab2735)

Overview

  • Product nameAnti-O-Linked N-Acetylglucosamine antibody [HGAC85] - ChIP Grade
    See all O-Linked N-Acetylglucosamine primary antibodies
  • Description
    Mouse monoclonal [HGAC85] to O-Linked N-Acetylglucosamine - ChIP Grade
  • Tested applicationsSuitable for: ChIP, IHC, ELISA, ICC/IF, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Rabbit, Cow
  • Immunogen

    Heat killed, pepsin treated group A streptococci.

  • General notesDO NOT USE WITH DILUENTS CONTAINING GLYCOSYLATED PROTEINS.

Properties

Applications

Our Abpromise guarantee covers the use of ab2735 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ChIP Use at an assay dependent concentration.
IHC Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
ICC/IF Use a concentration of 10 µg/ml.
IP Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml.

This antibody detects several proteins representing O-GlcNAc glycoproteins.

Target

  • RelevanceMany cellular proteins, including nuclear pore, oncogene, cytoskeletal, heat shock, viral and transcription regulatory proteins contain single O-linked N-acetylglucosamine (O-GlcNAc) residues attached to serine or threonine residues. It has been observed that O-GlcNAc glycosylated proteins tend to be under phosphorylated relative to unglycosylated proteins and that O-GlcNAc bearing proteins tend to be found in multimeric complexes. This has led to the suggestion that O-GlcNAc glycosylation may obscure phosphorylation sites and acts as a signaling mechanism or mediator of signaling.

Anti-O-Linked N-Acetylglucosamine antibody [HGAC85] - ChIP Grade images

  • Fluorescence analysis of a synthetic glycopolymer, staining O-Linked N-Acetylglucosamine with ab2735.

    A polydimethylsiloxane (PDMS) stamp was used for microcontact printing. The stamp was inked with a solution of glycopolymer (10 mM) and pressed into contact with freshly cleaned gold. The gold was then washed with absolute ethanol and dried with argon prior to incubating with primary antibody for 30 min followed by a AlexaFluor®568-conjugated goat-anti mouse secondary antibody for 30 min.
  • Anti-O-Linked N-Acetylglucosamine antibody [HGAC85] - ChIP Grade (ab2735) at 1/1000 dilution + Mouse liver tissue lysate - whole at 40 µg

    Secondary
    HRP-conjugated donkey anti-rabbit IgG polyclonal at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 28,54 kDa (why is the actual band size different from the predicted?)


    Exposure time : 1 minute

    This image is courtesy of an anonymous Abreview

    See Abreview

References for Anti-O-Linked N-Acetylglucosamine antibody [HGAC85] - ChIP Grade (ab2735)

This product has been referenced in:
  • Macauley MS  et al. Metabolism of vertebrate amino sugars with N-glycolyl groups: intracellular ß-O-linked N-glycolylglucosamine (GlcNGc), UDP-GlcNGc, and the biochemical and structural rationale for the substrate tolerance of ß-O-linked ß-N-acetylglucosaminidase. J Biol Chem 287:28882-97 (2012). WB ; Mouse . Read more (PubMed: 22692202) »
  • Vázquez-Dorbatt V  et al. Synthesis of a pyridyl disulfide end-functionalized glycopolymer for conjugation to biomolecules and patterning on gold surfaces. Biomacromolecules 10:2207-12 (2009). Small Molecule Fluorescent Optical Detection . Read more (PubMed: 19606855) »

See all 7 Publications for this product

Product Wall

Application Western blot
Loading amount 40 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Tissue lysate - whole (Liver)
Specification Liver
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Jun 02 2014

Application Western blot
Loading amount 10 µg
Gel Running Conditions Reduced Denaturing (10%)
Sample Human Cell lysate - whole cell (HepG2)
Specification HepG2
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
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Submitted Dec 10 2013

You are correct that the uniprot link in the previous email (Uniprot: http://www.uniprot.org/uniprot/Q6ZMB0http://www.uniprot.org/uniprot/Q6ZMB0) is for the core 3 synthase, which is B3GNT6. Inferring from the alternative names for this synthase (Beta-...

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I can confirm that the B3GNT6 antibodies, http://www.abcam.com/b3gnt6-antibody-ab103239.html& http://www.abcam.com/b3gnt6-antibody-ab168600.html, will recognize the transferase that synthesizes the core 3 structure of the O-glycan. We also curren...

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Thank you for your enquiry. I would check Sigma, I think that they probably carry it. If you have any further questions, let me know.

Thank you for your enquiry. Yes, if you have glycosylated proteins in your buffers (ie non-fat dry milk) that would bind the antibody. Therefore we would suggest glycosylated free BSA for your buffers. We regret to inform you that we don't have contro...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"