Proteasome Activity Assay试剂盒(ab107921)

Abreviews (1)Q&A (22)Specific References (9)

概述

  • 产品名称
    Proteasome Activity Assay试剂盒
  • 样品类型
    Cell Lysate
  • 检测类型
    Enzyme activity
  • 检测时间
    1h 00m
  • 种属反应性
    与反应: Human
  • 产品概述

    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.

  • 说明

    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

  • 经测试应用
    适用于: Functional Studiesmore details

性能

应用

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.

应用 Ab评论 说明
Functional Studies Use at an assay dependent dilution.

图片

  • Functional Studies - Proteasome Activity Assay Kit (ab107921)
    Functional Studies - Proteasome Activity Assay Kit (ab107921)

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

  • Functional Studies - Proteasome Activity Assay Kit (ab107921)
    Functional Studies - Proteasome Activity Assay Kit (ab107921)

    Proteasome Activity measured in Jurkat lysate

  • Functional Studies - Proteasome Activity Assay Kit (ab107921)
    Functional Studies - Proteasome Activity Assay Kit (ab107921)

    Proteasome Activity measured in HeLa lysate

  • Functional Studies - Proteasome Activity Assay Kit (ab107921)
    Functional Studies - Proteasome Activity Assay Kit (ab107921)

    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.

实验方案

文献

This product has been referenced in:
  • Sambri I  et al. Lysosomal dysfunction disrupts presynaptic maintenance and restoration of presynaptic function prevents neurodegeneration in lysosomal storage diseases. EMBO Mol Med 9:112-132 (2017). Read more (PubMed: 27881461) »
  • Mi X  et al. Xanthohumol induces paraptosis of leukemia cells through p38 mitogen activated protein kinase signaling pathway. Oncotarget 8:31297-31304 (2017). Read more (PubMed: 28415750) »

See all 9 Publications for this product

发表研究结果有使用 ab107921？请让我们知道，以便我们可以引用本数据表中的参考文章。

客户评价及客户问答

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23 reviews or Q&A

10
Votes: Highest First

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.
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Username

Mr. Baptiste Jude

Verified customer

提交于 Oct 07 2016

How should samples be prepared?

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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I would like to know if RIPA buffer would be appropriate to use in the homogeneization step.

Thank you for your inquiry.

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

Will you confirm that the protease inhibitors NaF, PMSF, Leupeptin, Aprotinin, Bestatin, E-64, AEBSF and Pepstatin which are added to the RIPA buffer won't interfere with this assay, please?









<...

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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.

I am a little confused about what kind of plate to use. The protocol section recommends a white opaque plate, but the troubleshoot section suggests a black plate for fluorescence. If the proteasome assay is a fluorescence based assay, what would be the...

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

Dear technical specialist,
Regarding to the product of Proteasome-Activity-Assay-Kit-ab107921, I
would ask you a few questions here:
1. Do I need to make the standard curve everytime when I detect the samples?
2. Are cells pellets froze...

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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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HI

I need to choose two time points, T1 and T2. Please see the attached file from my new run. Most samples were run 1:5, a couple 1:10 because I didn't have enough sample.
So I've chosen two time points of each sample, I've marked them ...

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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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Should I dilute the samples more, would that be useful? Now the curves max out very early after just few minutes. I used 50ul cell lysates, which is 1:2 dilution. Maybe 1:10 would be better?

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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I have trouble analysing the results. I don’t know how to choose T1 and T2. Could you please look at my plots and give some advice.

Am I supposed to add the inhibitor to one of the standard wells as well?

What should I use as blank fo...

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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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Yet another question regarding this kit (I don’t think the protocol is very clear). What should I use as blank for the measurements since the machine then calculates values based on blank correction. Should I use 10ul of the cell lysate buffer +9...

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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"

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