Synthetic Human Ubiquitin vinyl methyl ester (HA-tag) protein (ab206172)

Description

  • Product name

    Synthetic Human Ubiquitin vinyl methyl ester (HA-tag) protein
  • Expression system

    Synthetic
  • Accession

  • Protein length

    Full length protein
  • Animal free

    No
  • Nature

    Synthetic
    • Species

      Human
    • Predicted molecular weight

      10 kDa including tags
    • Amino acids

      1 to 76
    • Additional sequence information

      N-terminal HA tag (YPYDVPDYA).

Specifications

Our Abpromise guarantee covers the use of ab206172 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

  • Applications

    HPLC

    SDS-PAGE

    Mass Spectrometry

  • Form

    Lyophilised
  • Additional notes

    ab206172 is a potent, irreversible and specific inhibitor of deubiquitinating enzymes (DUBs) that is prepared by chemical synthesis. It is N-terminally tagged with an HA-tag (YPYDVPDYA), which allows for sensitive identification or purification of DUBs since it is specifically recognized by anti-HA antibodies. The HA tag is separated from the Ub N-terminus by two aminohexanoic acid (Ahx) linkers for efficient recognition of the HA tag.

  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped at 4°C. Store at -20°C. Avoid freeze / thaw cycle.

    To prepare the sample, dissolve the powder in DMSO (e.g. add 1.25 µl DMSO to 25 µg powder to get 20 mg/ml or 2.25 mM DMSO stock) and then slowly add this DMSO stock to the required buffer (please note the order of addition).

  • Reconstitution
    Dissolve the powder in DMSO (e.g. add 1.25 µl DMSO to 25 µg powder to get 20 mg/ml or 2.25 mM DMSO stock) and add this DMSO stock to the required buffer (please note the order of addition). Store reconstituted solution at -80°C.

General Info

  • Relevance

    The ubiquitin cascade in its many forms includes a large family of cysteine proteases known as deubiquitinylating enzymes (DUBs). Many members are assigned to this enzyme class by sequence similarity but with little evidence for specific biological activity. HA-tagged ubiquitin molecules derivatised by chemical ligation at the C-terminus with thiol-reactive functional groups can be used as potent, irreversible inhibitors of DUBs through their covalent modification of the active site and, consequently, can act as specific probes for enzymes with DUB activity. Such probes have been utilized for the identification of DUBs and associated components from cell lysates by proteomic methods, as well as rapid demonstration of enzymatic activity for gene products whose functions were inferred from primary structure. The nature of the electrophilic group used as the C-terminal derivative, for example vinyl sulfone (-VS), vinyl methylester (-VME), 2-bromoethyl (-Br) or 2-chloroethyl (-Cl), may determine the subset of DUB enzymes detected. The HA peptide sequence (YPYDVPDYA), derived from the influenza hemagglutinin protein, facilitates sensitive identification or purification of HA-Ub-VME modified DUBs through recognition by HA-reactive antibodies in free or agarose-immobilised.

Images

  • Coomassie-stained SDS-PAGE analysis of ab206172.

    Lane 1: ab206172
    Lane 2: Ubiquitin

References

ab206172 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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