Anti-O-Linked N-Acetylglucosamine antibody [RL2] (Alexa Fluor® 488) (ab201993)

Mouse monoclonal O-Linked N-Acetylglucosamine antibody [RL2] conjugated to Alexa Fluor® 488. Validated in Flow Cyt, ICC/IF and tested in Human.

Overview

  • Product name

    Anti-O-Linked N-Acetylglucosamine antibody [RL2] (Alexa Fluor® 488)
    See all O-Linked N-Acetylglucosamine primary antibodies
  • Description

    Mouse monoclonal [RL2] to O-Linked N-Acetylglucosamine (Alexa Fluor® 488)
  • Host species

    Mouse
  • Conjugation

    Alexa Fluor® 488. Ex: 495nm, Em: 519nm
  • Tested applications

    Suitable for: Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Positive control

    • ICC/IF: MCF7 cells. Flow Cyt: MCF7 cells.
  • General notes

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped 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 buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 30% Glycerol, 1% BSA
  • Concentration information loading...
  • Purity

    IgG fraction
  • Clonality

    Monoclonal
  • Clone number

    RL2
  • Isotype

    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab201993 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
Flow Cyt 1/500.
ICC/IF 1/200.

This product gave a positive signal in MCF7 cells fixed with 4% formaldehyde (10 min) and 100% methanol (5 min).

Target

  • Relevance

    Many 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.

Images

  • Overlay histogram showing MCF7 cells stained with ab201993 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab201993, 1/500 dilution) for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 (monoclonal) Alexa Fluor® 488 (ab171463) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in MCF7 cells fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • ab201993 staining O-Linked N-Acetylglucosamine in MCF7 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab201993 at 1/200 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This product also gave a positive signal under the same testing conditions in MCF7 cells fixed with 100% methanol (5min).

References

ab201993 has not yet been referenced specifically in any publications.

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Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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