Product nameAnti-O-Linked N-Acetylglucosamine antibody [RL2] (HRP)
See all O-Linked N-Acetylglucosamine primary antibodies
DescriptionMouse monoclonal [RL2] to O-Linked N-Acetylglucosamine (HRP)
Tested applicationsSuitable for: IHC-P, WBmore details
Species reactivityReacts with: Human
- IHC-P: Normal human colon tissue. WB: Jurkat treated with 0 uM PugNAc and Jurkat treated with 4 mM glucosamine and 50 uM PugNAc (3 hours).
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. Store In the Dark.
Storage bufferpH: 7.40
Preservative: 0.1% Proclin
Constituents: PBS, 30% Glycerol, 1% BSA
Concentration information loading...
Our Abpromise guarantee covers the use of ab201995 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
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.
All lanes : Anti-O-Linked N-Acetylglucosamine antibody [RL2] (HRP) (ab201995) at 1/1000 dilution
Lane 1 : Jurkat treated with 0 uM PugNAc
Lane 2 : Jurkat treated with 4 mM glucosamine and 50 uM PugNAc (3 hours)
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 100,65 kDa why is the actual band size different from the predicted?
Exposure time: 2 minutes
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab201995 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.
IHC image of O-linked N-Acetylglucosamine staining in a section of formalin-fixed paraffin-embedded normal human colon tissue*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab201995, 1/100 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.
ab201995 has not yet been referenced specifically in any publications.