Recombinant
RabMAb

Recombinant Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (Alexa Fluor® 647) (ab192489)

Overview

  • Product name

    Anti-Ubiquitin (linkage-specific K63) antibody [EPR8590-448] (Alexa Fluor® 647)
    See all Ubiquitin primary antibodies
  • Description

    Rabbit monoclonal [EPR8590-448] to Ubiquitin (linkage-specific K63) (Alexa Fluor® 647)
  • Host species

    Rabbit
  • Conjugation

    Alexa Fluor® 647. Ex: 652nm, Em: 668nm
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human Ubiquitin aa 50-150 (Cysteine residue). The exact sequence is proprietary. (K63-linkage specific polyubiquitin).
    Database link: P0CG48

  • Positive control

    • ICC/IF - HeLa, Flow Cyt - HeLa
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab192489 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 Use 2µl for 106 cells.

ab199093 - Rabbit monoclonal IgG (Alexa Fluor® 647), is suitable for use as an isotype control with this antibody.

 

ICC/IF 1/50.

Target

  • Relevance

    Function: Ubiquitin exists either covalently attached to another protein, or free (unanchored). When covalently bound, it is conjugated to target proteins via an isopeptide bond either as a monomer (monoubiquitin), a polymer linked via different Lys residues of the ubiquitin (polyubiquitin chains) or a linear polymer linked via the initiator Met of the ubiquitin (linear polyubiquitin chains). Polyubiquitin chains, when attached to a target protein, have different functions depending on the Lys residue of the ubiquitin that is linked: Lys-6-linked may be involved in DNA repair; Lys-11-linked is involved in ERAD (endoplasmic reticulum-associated degradation) and in cell-cycle regulation; Lys-29-linked is involved in lysosomal degradation; Lys-33-linked is involved in kinase modification; Lys-48-linked is involved in protein degradation via the proteasome; Lys-63-linked is involved in endocytosis, DNA-damage responses as well as in signaling processes leading to activation of the transcription factor NF-kappa-B. Linear polymer chains formed via attachment by the initiator Met lead to cell signaling. Ubiquitin is usually conjugated to Lys residues of target proteins, however, in rare cases, conjugation to Cys or Ser residues has been observed. When polyubiquitin is free (unanchored-polyubiquitin), it also has distinct roles, such as in activation of protein kinases, and in signaling. Similarity: Belongs to the ubiquitin family. Contains 3 ubiquitin-like domains.
  • Cellular localization

    Cell Membrane, Cytoplasmic and Nuclear
  • Database links

  • Alternative names

    • Epididymis secretory protein Li 50 antibody
    • FLJ25987 antibody
    • HEL S 50 antibody
    • MGC8385 antibody
    • Polyubiquitin B antibody
    • RPS 27A antibody
    • RPS27A antibody
    • UBA 52 antibody
    • UBA 80 antibody
    • UBA52 antibody
    • UBA80 antibody
    • UBB antibody
    • UBB_HUMAN antibody
    • UBC antibody
    • UBCEP 1 antibody
    • UBCEP 2 antibody
    • UBCEP1 antibody
    • UBCEP2 antibody
    • Ubiquitin antibody
    • Ubiquitin B antibody
    see all

Images

  • ab192489 staining Ubiquitin (linkage-specific K63) in HeLa cells. The cells were fixed with 100% methanol (10 min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Triton X-100 for 1hr. The cells were then incubated with ab192489 at a working dilution of 1 in 50 (shown in red) and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin) at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1hr with an AlexaFluor® 488 Goat anti-mouse IgG (H&L - preadsorbed) secondary (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    This product gave a positive signal in 4% formaldehyde (10 min) fixed HeLa cells under the same testing conditions.

    Image was taken with a Confocal microscope (Leica micro-systems, TCS SP8).

  • Overlay histogram showing HeLa cells stained with ab192489 (red line). The cells were fixed with 4% formaldehyde (10 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 (ab192489, 2μl/1x106 cells) for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (Alexa Fluor® 647) (2μl/1x106 cells) for 30 min at 22ºC. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 25mW red solid state diode laser (635nm) and 675/30 bandpass filter.

References

ab192489 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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