Recombinant UBE2I / UBC9 protein (ab3803)

概述

描述

  • 性质Recombinant
  • 来源Escherichia coli

技术指标

Our Abpromise guarantee covers the use of ab3803 in the following tested applications.

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

  • 生物活性The final fraction of enzyme contains a single polypeptide band of 18 kDa.
  • 应用

    Western blot

    Functional Studies

  • 形式Liquid
  • 补充说明


    This product can be used as part of an assay for sumoylation activity. Human Aos 1 + Uba 2 (ab3804), Ubc 9 (ab3803) and Sumo 1 (ab3801) can be used to promote in vitro sumoylation of a sumoylation marker (human Topoisomerase I protein fragment) (ab3828). The reaction products can be detected using our Sumo 1 (ab3819 and ab3824) and Topoisomerase I (ab3825) antibodies. Sumoylation assays are carried out in a final volume of 20µl in reaction conditions (20 mM Hepes pH 7.5, 5mM MgCl2, 2mM ATP). Sumoylation Protocol: 1. Prepare a suitable purified substrate protein. (For the control, use 2µl Topoisomerase I marker for each reaction.) 2. In each reaction, add 4µl E2 to substrate first, then 2µl Sumo 1, 2µl 10x reaction buffer, 2µl E1. Finally, add H2O to bring up to 20µl. We would recommend adding fresh 2mM ATP to be sure that sufficient energy is supplied. 3. The best reaction concentration of proteins is as following: Aos 1 + Uba 2: 7.5µg/ml. Ubc 9: 50µg/ml. SUMO 1: 50µg/ml. For the control assay we recommend running the assay at 37ºC for 30-60 minutes. 4. Detect the reaction products by Western blot using a suitable antibody. For the control reaction use 1/1000 dilution of the supplied Topoisomerase I antibody. Four sumoylated bands should be seen on the gel for the control reaction. This assay has been shown to work with crude extracts. Be aware that Uba 2 contains his-rich regions which might cross-react with antibodies against the 6x-His epitope tag. During western analysis with anti-6x-His antibodies, Uba 2 at 80 kDa might be shown.

    The final fraction of enzyme contains a single polypeptide band of 18 kDa.
  • Concentration information loading...

制备和贮存

  • 稳定性和存储

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

    10 mM Tris-Cl, pH 7.5, 100 mM NaCl, 100 mM imidazole, 0.5 mM PMSF, 1 mM DTT, and 10 % glycerol

常规信息

  • 别名
    • C358B7.1
    • p18
    • SUMO 1 protein ligase
    • SUMO conjugating enzyme UBC9
    • SUMO-conjugating enzyme UBC9
    • SUMO-protein ligase
    • SUMO1 protein ligase
    • UBC9
    • UBC9_HUMAN
    • UBCE9
    • Ube2i
    • Ubiquitin carrier protein
    • Ubiquitin carrier protein 9
    • Ubiquitin carrier protein I
    • Ubiquitin conjugating enzyme 9
    • Ubiquitin conjugating enzyme E2I (homologous to yeast UBC9)
    • Ubiquitin conjugating enzyme E2I (UBC9 homolog, yeast)
    • Ubiquitin conjugating enzyme UbcE2A
    • Ubiquitin like protein SUMO 1 conjugating enzyme
    • Ubiquitin protein ligase E2I
    • Ubiquitin-conjugating enzyme E2 I
    • Ubiquitin-protein ligase I
    see all
  • 功能Accepts the ubiquitin-like proteins SUMO1, SUMO2, SUMO3 and SUMO4 from the UBLE1A-UBLE1B E1 complex and catalyzes their covalent attachment to other proteins with the help of an E3 ligase such as RANBP2 or CBX4. Necessary for sumoylation of FOXL2 and KAT5. Essential for nuclear architecture and chromosome segregation.
  • 组织特异性Expressed in heart, skeletal muscle, pancreas, kidney, liver, lung, placenta and brain. Also expressed in testis and thymus.
  • 通路Protein modification; protein sumoylation.
  • 序列相似性Belongs to the ubiquitin-conjugating enzyme family.
  • 细胞定位Nucleus. Cytoplasm. Mainly nuclear. In spermatocytes, localizes in synaptonemal complexes. Recruited by BCL11A into the nuclear body.
  • Information by UniProt

Recombinant UBE2I / UBC9 protein 图像

  • Left: Human topo I (S35Met-labeled) control.

    Right: Sumoylated human topo I (S35Met-labeled)

    (7.5 µg/ml E1, 50 µg/ml E2, 50 µg/ml sumo1, 37ºC 30min).

  • Extracts prepared from CEF cells not transfected (-) or transfected (+) with wild-type Src were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to nitrocellulose. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then were incubated with 1.0 µg/mL anti-Src pan antibody. After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG alkaline phosphatase and the signal was detected using the Tropix WesternStar method.

Recombinant UBE2I / UBC9 protein (ab3803)参考文献

ab3803 has not yet been referenced specifically in any publications.

Product Wall

Thank you for getting back to me. According to the details that I have Ubc9 protein (ab3803) is not epitope tagged. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Thank you for your enquiry. The Uba 2 protein ab3803 is not tagged. However, it does contain his-rich regions which might cross-react with antibodies against the 6x-His epitope tag. According to our datasheet during western analysis with anti-6x-His...

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Thank you for your enquiry. Ab3803 is Ubc 9 protein, not an antibody, and has only been tested so far in Western blotting and Functional Studies. If you have any more questions, please do contact us again.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"