Anti-O-Linked N-Acetylglucosamine antibody [RL2] (ab2739)
Overview
- Product nameAnti-O-Linked N-Acetylglucosamine antibody [RL2]
See all O-Linked N-Acetylglucosamine primary antibodies - DescriptionMouse monoclonal [RL2] to O-Linked N-Acetylglucosamine
- SpecificityDetects nuclear pore complex (NPC), cytoplasmic and intranuclear O-linked glycoproteins from human, mouse, and rat tissues.
- Tested applicationsICC/IF, IHC-Fr, ChIP/Chip, Dot Blot, WB, IPmore 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 instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
- Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Some batches may contain 0.4M arginine. -
Concentration information loading... - PurityIgG fraction
- Clonality Monoclonal
- Clone numberRL2
- IsotypeIgG1
- Research Areas
Applications
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.
| Application | Abreviews | Notes |
|---|---|---|
| ICC/IF | Use a concentration of 5 - 10 µg/ml. | |
| IHC-Fr | Use at an assay dependent concentration. PubMed: 23734074 | |
| ChIP/Chip | Use at an assay dependent concentration. PubMed: 20368426 | |
| Dot Blot | 1/800. | |
| WB | Use a concentration of 1 µg/ml. | |
| IP | Use at an assay dependent concentration. |
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.
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Alternative names
- O-GlcNAc antibody
Anti-O-Linked N-Acetylglucosamine antibody [RL2] images
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![Immunocytochemistry/ Immunofluorescence - O-Linked N-Acetylglucosamine antibody [RL2] (ab2739)](http://a.abcam.com/ps/datasheet/images/2/ab2739/O-Linked-N-Acetylglucosamine-Primary-antibodies-ab2739-1.jpeg)
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 - Anti-O-Linked N-Acetylglucosamine antibody [RL2] (ab2739)](http://a.abcam.com/ps/products/2/ab2739/Images/ab2739-232732-ab2739wb.jpg)
Western blot - Anti-O-Linked N-Acetylglucosamine antibody [RL2] (ab2739)This image is courtesy of an anonymous AbreviewAll lanes : Anti-O-Linked N-Acetylglucosamine antibody [RL2] (ab2739) at 1/3000 dilution
Lane 1 : Human neuroblastoma (SH-SY5Y) whole cell lysate - treated with 50µM z-Pugnac for 24 hours
Lane 2 : Human neuroblastoma (SH-SY5Y) whole cell lysate - untreated
Lysates/proteins at 20 µg per lane.
Secondary
HRP-conjugated horse anti-mouse IgG polyclonal
developed using the ECL technique
Performed under reducing conditions.
Exposure time : 30 secondsThis image is courtesy of an anonymous Abreview
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![Western blot - O-Linked N-Acetylglucosamine antibody [RL2] (ab2739)](http://a.abcam.com/ps/datasheet/images/2/ab2739/O-Linked-N-Acetylglucosamine-Primary-antibodies-ab2739-4.jpg)
Western blot - O-Linked N-Acetylglucosamine antibody [RL2] (ab2739)Anti-O-Linked N-Acetylglucosamine antibody [RL2] (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 minuteThe antibody was tested against the immunogen (isolated rat liver nuclear envelopes, which contain 8 O-linked glycoproteins in the nuclear pore complex). -
![Western blot - Anti-O-Linked N-Acetylglucosamine antibody [RL2] (ab2739)](http://a.abcam.com/ps/products/2/ab2739/Images/ab2739-232645-WBAP10859411.jpg)
Western blot - Anti-O-Linked N-Acetylglucosamine antibody [RL2] (ab2739)All lanes : Anti-O-Linked N-Acetylglucosamine antibody [RL2] (ab2739) at 1 µg/ml
Lane 1 : Jurkat cells treated with 0 uM PugNAc
Lane 2 : Jurkat cells treated with 50 uM PugNAc (3 hours)
Lane 3 : Jurkat cells treated with 0 uM PugNAc
Lane 4 : Jurkat cells treated with 4 mM glucosamine and 50 uM PugNAc (3 hours)
Lysates/proteins at 20 µg per lane.
Secondary
Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 dilution
developed using the ECL technique
Performed under reducing conditions.
Exposure time : 12 minutesJurkat cells were treated with either 50 uM PugNAc (ab144670) or 4 mM glucosamine + 50 uM PugNAc (ab144670) for three hours prior to harvest to stimulate O-linked glycosylation. The expected increase in glycosylation is observed in the treated lanes 2 & 4.
References for Anti-O-Linked N-Acetylglucosamine antibody [RL2] (ab2739)
This product has been referenced in:
- Chu CS et al. O-GlcNAcylation regulates EZH2 protein stability and function. Proc Natl Acad Sci U S A 111:1355-60 (2014). Read more (PubMed: 24474760) »
- Zhang Q et al. Differential regulation of the ten-eleven translocation (TET) family of dioxygenases by O-linked ß-N-acetylglucosamine transferase (OGT). J Biol Chem 289:5986-96 (2014). Read more (PubMed: 24394411) »
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"