Anti-O-Linked N-Acetylglucosamine antibody [RL2] - ChIP Grade (ab2739)
- Product nameAnti-O-Linked N-Acetylglucosamine antibody [RL2] - ChIP GradeSee all O-Linked N-Acetylglucosamine primary antibodies ...
- DescriptionMouse 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
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).
- 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
- Research Areas
Our Abpromise guarantee covers the use of ab2739 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||ICC/IF: Use a concentration of 5 - 10 µg/ml.|
|ChIP/Chip||ChIP/Chip: Use at an assay dependent dilution. PubMed: 20368426|
|Dot Blot||Dot: 1/800.|
|WB||WB: Use a concentration of 1 µg/ml.|
|IP||IP: Use at an assay dependent dilution.|
- 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.
- O-GlcNAc antibody
Anti-O-Linked N-Acetylglucosamine antibody [RL2] - ChIP Grade images
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.
Anti-O-Linked N-Acetylglucosamine antibody [RL2] - ChIP Grade (ab2739) at 1 µg/ml + Rat Liver Nuclear Envelope at 10 µg
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
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.
References for Anti-O-Linked N-Acetylglucosamine antibody [RL2] - ChIP Grade (ab2739)
This product has been referenced in:
- Fong JJ et al. ß-N-Acetylglucosamine (O-GlcNAc) Is a Novel Regulator of Mitosis-specific Phosphorylations on Histone H3. J Biol Chem 287:12195-203 (2012). WB . Read more (PubMed: 22371497) »
- Riedl E et al. Carnosine prevents apoptosis of glomerular cells and podocyte loss in STZ diabetic rats. Cell Physiol Biochem 28:279-88 (2011). Read more (PubMed: 21865735) »