Alexa Fluor® 594 anti-O-GlcNAc Antibody

Pricing & Availability
Clone
CTD110.6 (See other available formats)
Other Names
O-Linked N-Acetylglucosamine
Isotype
Mouse IgM, κ
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Product Citations
publications
CTD110.6_A594_OGlcNAc_Antibody_IHCP_100616
Human paraffin-embedded intestine tissue slices were prepared with a standard protocol of deparaffination and rehydration. Antigen retrieval was done with Tris-Buffered Saline 20X (1.0M, pH7.4) at 95°C for 40 minutes. Tissue was washed with PBS/ 0.05% Tween20 twice for five minutes and blocked with 5% FBS and 0.2% Gelatin for 30 minutes. Then, the tissue was stained with 5 µg/ml of anti-O-GlcNAc (clone CTD110.6) Alexa Fluor® 594 (red) at 4°C overnight. Nuclei were counterstained with DAPI (blue). The image was captured with a 10X objective.
  • CTD110.6_A594_OGlcNAc_Antibody_IHCP_100616
    Human paraffin-embedded intestine tissue slices were prepared with a standard protocol of deparaffination and rehydration. Antigen retrieval was done with Tris-Buffered Saline 20X (1.0M, pH7.4) at 95°C for 40 minutes. Tissue was washed with PBS/ 0.05% Tween20 twice for five minutes and blocked with 5% FBS and 0.2% Gelatin for 30 minutes. Then, the tissue was stained with 5 µg/ml of anti-O-GlcNAc (clone CTD110.6) Alexa Fluor® 594 (red) at 4°C overnight. Nuclei were counterstained with DAPI (blue). The image was captured with a 10X objective.
See Alexa Fluor® 594 spectral data
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838006 100 µg $345
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Description

β-O-linked N-acetylglucosamine (O-GlcNAc) is an abundant post-translational modification occuring on serine and threonine residues. The modification has been detected on a variety of nuclear and cytoplasmic proteins involved in diverse cellular functions. Studies to elucidate the role of O-GlcNAc-ylation have been hampered by the lack of a sensitive, specific assay to identify or purify proteins bearing O-GlcNAc.

Product Details
Technical data sheet

Product Details

Reactivity
All Species
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
The antibody was raised against a synthetic peptide containing Serine-O-GlcNAc.
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 594 under optimal conditions. The solution is free of unconjugated Alexa Fluor® 594.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

IF - Quality tested
IHC-P - Validated

Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescence staining. For immunofluorescence microscopy, a concentration range of 1.25 - 5.0 μg/ml is recommended. For immunohistochemical staining on formalin-fixed paraffin-embedded tissue sections, the suggested use of this reagent is 5.0 - 10 µg/ml. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 594 has an excitation maximum of 590 nm, and a maximum emission of 617 nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Application Notes

This antibody detects Ser-O-GlcNAc and Thr-O-GlcNAc but shows no cross-reactivity with peptide determinants or other closely-related carbohydrate antigens. Interaction between the antibody and the Ser- or Thr-O-GlcNAc epitope can be blocked by competition with free GlcNAc. Milk proteins may cross-react with the antibody therefore we recommend using BSA or goat serum as a blocker.

Additional reported applications (for the relevant formats) include: ELISA2, immunocytochemistry7, immunoprecipitation2,5,6, and Western blotting3,4,7.

Application References

(PubMed link indicates BioLegend citation)
  1. Comer FI, et al. 2001. Anal. Biochem. 293:169.
  2. Hart GW, et al. 1995. Adv. Exp. Med. Biol. 376:115. (ELISA, WB, IP)
  3. Anthonisen EH, et al. 2010. J. Biol. Chem. 285:1607. (WB)
  4. Dias WB, et al. 2009. J. Biol. Chem. 32:21327. (WB)
  5. Wang Z, et al. 2007. Mol. Cell Proteomics 8:1365. (IP)
  6. Kim SJ, et al. 2003. J. Biol. Chem. 43:41890. (IP)
  7. Kneass ZT, Marchase RB. 2004. J. Biol. Chem. 44:45759. (WB, ICC, IF)
RRID
AB_2632636 (BioLegend Cat. No. 838006)

Antigen Details

Structure
1046 amino acids with a molecular weight of 117 kD.
Distribution

Cytosol, mitochondrion, nucleus, plasma membrane, and cytoskeleton.

Function
Catalyzes the transfer of a single N-acetylglucosamine from UDP-GlcNAc to a serine or threonine residue in cytoplasmic and nuclear proteins resulting in their modification with a beta-linked N-acetylglucosamine (O-GlcNAc), Glycosylates a large and diverse number of proteins including histone H2B, AKT1, EZH2, PFKL, KMT2E/MLL5, MAPT/TAU and HCFC1, Can regulate their cellular processes via cross-talk between glycosylation and phosphorylation or by affecting proteolytic processing.
Biology Area
Cell Biology, Transcription Factors
Gene ID
8473 View all products for this Gene ID
UniProt
View information about O-GlcNAc on UniProt.org

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Go To Top Version: 3    Revision Date: 11/08/2016

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