Anti-O-Linked N-Acetylglucosamine antibody [RL2] - ChIP Grade (ab2739)

Overview

• Product nameAnti-O-Linked N-Acetylglucosamine antibody [RL2] - ChIP GradeSee all O-Linked N-Acetylglucosamine primary antibodies ...
• Description
  Mouse monoclonal [RL2] to O-Linked N-Acetylglucosamine - ChIP Grade
• SpecificityDetects nuclear pore complex (NPC), cytoplasmic and intranuclear O-linked glycoproteins from human, mouse, and rat tissues.
• Tested applicationsICC/IF, ChIP/Chip, Dot Blot, WB, IP more details
• Immunogen

  Pore complex-lamina fraction purified from rat liver nuclear envelopes.

• General notesWe can conjugate this antibody to FITC for you (please see ab150247 for details).

Properties

• FormLiquid
• Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
• Storage bufferPBS with 0.05% sodium azide
• Concentration information loading...
• PurityIgG fraction
• Clonality Monoclonal
• Clone numberRL2
• IsotypeIgG1
• Research Areas
  • Signal Transduction
  • Protein Trafficking
  • Nuclear Import / Export

  • Epigenetics and Nuclear Signaling
  • ChIP'ing antibodies
  • ChIP'ing antibodies

  • Neuroscience
  • Neurology process
  • Neurodegenerative disease
  • Alzheimer's disease
  • Other

  • Cell Biology
  • Other Antibodies
  • Other Antibodies

Applications

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.

• Alternative names
  O-GlcNAc antibody

Anti-O-Linked N-Acetylglucosamine antibody [RL2] - ChIP Grade images

• 

  Immunocytochemistry/ Immunofluorescence - O-Linked N-Acetylglucosamine antibody [RL2] (ab2739)
  ICC/IF image of ab2739 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2739, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HEK293, HepG2, and MCF-7 cells at 10µg/ml, and in 100% Methanol fixed (5 min) HeLa, Hek293, HepG2, and MCF-7 cells at 5µg/ml.
• 

  Western blot - O-Linked N-Acetylglucosamine antibody [RL2] (ab2739)
  Anti-O-Linked N-Acetylglucosamine antibody [RL2] - ChIP Grade (ab2739) at 1 µg/ml + Rat Liver Nuclear Envelope at 10 µg
  
  Secondary
  Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
  developed using the ECL technique
  
  Performed under reducing conditions.
  
  Exposure time : 1 minute
• 

  Immunocytochemistry/ Immunofluorescence-Anti-O-Linked N-Acetylglucosamine antibody [RL2] - ChIP Grade(ab2739)
  ab2739 staining O-linked N-Acetylglucosamine in HeLa cells treated with tunicamycin from Streptomyces sp (ab120296), by ICC/IF. Decrease in O-linked N-Acetylglucosamine expression correlates with increased concentration of tunicamycin from Streptomyces sp, as described in literature.
  The cells were incubated at 37°C for 24h in media containing different concentrations of ab120296 (tunicamycin from Streptomyces sp) in DMSO, fixed with 100% methanol for 5 minutes at -20°C and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab2739 (10 µg/ml) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-mouse polyclonal antibody (ab96879) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.