製品の概要

  • 製品名Proteasome Activity Assay Kit
  • テスト
    100 x 1 assay
  • サンプルの種類
    Cell Lysate
  • アッセイタイプEnzyme activity
  • 全工程の試験時間
    1h 00m
  • 種交差性
    交差種: Human
  • 製品の概要

    Proteasome Activity Assay Kit (ab107921) takes advantage of the chymotrypsin-like activity, utilizing an AMC-tagged peptide substrate (Proteasome Substrate (Succ-LLVY-AMC in DMSO), which releases free, highly fluorescent AMC in the presence of proteolytic activity. The kit also includes a positive control (Jurkat Cell lysate with significant proteasome activity) and a specific proteasome inhibitor MG-132 which suppresses all proteolytic activity due to proteasomes. This permits differentiation of proteasome activity from other protease activity which may be present in samples.
    Visit our FAQs page for tips and troubleshooting.

  • 特記事項

    Proteasomes are very large (20S, 26S) protein assemblies found in both the nucleus and cytoplasm of all eucaryotes (and in some procaryotes). They are responsible for the degradation and recycling of proteins which have been previously tagged with ubiquitin. Such tagged proteins are degraded into peptides approximately 7-8 amino acids long which are subsequently further degraded. The 20S assembly is the functional protease structure with chymotrypsin-like, trypsin-like and caspase-like protease activities

  • アプリケーション適用あり: Functional Studiesmore details

製品の特性

    アプリケーション

    Our Abpromise guarantee covers the use of ab107921 in the following tested applications.

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

    アプリケーション Abreviews 特記事項
    Functional Studies Use at an assay dependent dilution.

    Proteasome Activity Assay Kit 画像

    • Standard curve: mean of duplicates (+/- SD) with background reads subtracted

    • Proteasome Activity measured in Jurkat lysate

    • Proteasome Activity measured in HeLa lysate

    • Proteasome Activity measured in cell lysates showing activity (mU) per 1 mln cells.

      Samples with the concentration of 3.3e6 cells/mL were used. 50 μl of each sample was used.

    プロトコール

    Proteasome Activity Assay Kit (ab107921) 使用論文

    This product has been referenced in:

    See 1 Publication for this product

    Product Wall

    Proteasome activity in rat muscle

    Good Good 4/5 (Ease of Use)
    Abreviews
    Soleus muscle was used.10mg of muscle was homogenized in 100µl of 0.5% NP-40, and 20µl of sample was used in duplicate in the plate.
    Username

    Mr. Baptiste Jude

    Verified customer

    投稿 Oct 07 2016

    Start with 1 - 2 x 1e6 cells. Suspend the cell pellet in 500 μl (˜4 volumes) of the 0.5% NP40 solution on ice. Homogenize with a Dounce homogenizer (10 - 50 strokes) or equivalent on ice until efficient lysis is confirmed by viewing the cells under the...

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    Thank you for your inquiry.

    We do not recommend using samples prepared with RIPA buffer because the SDS tends to interfere with the enzyme.

    Addition of protease inhibitors for this assay (ab107921: Proteasome Activity Assay Kit) actually works well; since they will inhibit any proteases from degrading the proteins in the proteasome complex.


    Please use a white opaque plate. The troubleshooting guide (which recommends using a black plate for fluorescence) is just a general one.

    Thank you for contacting us.


    I am pleased to answer your questions regarding the Proteasome Activity Assay Kit (ab107921).

    1. I can confirm that it is necessary to create a new standard curve for every performed experiment. This ...

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    Thank you very much for your email.

    As my colleague is not in the office today, I have looked into your inquiry.

    I am happy to confirm that you can use any two time points as long as they are in the linear range. The difference betw...

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    The standard graph looks good. Based on that,you are right in saying that the samples can be diluted further maybe 1:5 this time for better time points.
    From this run, assuming that the X axis is time, I think maybe 8 and 10 (units unknown) would f...

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    Could you please send the standard curve for the attached plot?
    My colleague would be able to help with the T1 and T2 looking at the curve.

    Alternatively, this is the way you could decide the time points:
    It is essential to read R...

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    I just wanted to let you know that I am still working on your inquiry. I have contacted the originator of this product for the information you requested, and I am awaiting a reply.

    I am very sorry for the delay. I will forward to you the infor...

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