Cathepsin B Activity Assay试剂盒(Fluorometric) (ab65300)

概述

  • 产品名称Cathepsin B Activity Assay试剂盒(Fluorometric)
    参阅全部 Cathepsin B 试剂盒
  • 测试
    100 x 1 assay
  • 样品类型
    Tissue Extracts, Cell Lysate
  • 检测类型Quantitative
  • 检测时间
    2h 00m
  • 产品概述

    Cathepsin B Activity Assay kit (Fluorometric) (ab65300) is a fluorescence-based assay that utilizes the preferred cathepsin-B substrate sequence RR labeled with AFC (amino-4-trifluoromethyl coumarin). Cell lysates or other samples that contain cathepsin-B will cleave the synthetic substrate RR-AFC to release free AFC. The released AFC can easily be quantified using a fluorometer or fluorescence plate reader. The Cathepsin B assay is simple, straightforward, and can be adapted to 96-well plate assays. Assay conditions have been optimized to obtain the maximal activity.
    Visit our FAQs page for tips and troubleshooting.

  • 说明

    Apoptosis can be mediated by mechanisms other than the traditional caspase-mediated cleavage cascade. There is growing recognition that alternative proteolytic enzymes such as the lysosomal cathepsin proteases may initiate or propagate proapoptotic signals. Cathepsins are lysosomal enzymes that are also used as sensitive markers in various toxicological investigations.

  • 经测试应用适用于: Functional Studiesmore details

性能

  • 存放说明Store at -20°C. Please refer to protocols.
  • 组件 标识符 100 tests
    CB Cell Lysis Buffer WM 1 x 25ml
    CB Inhibitor (1mM) Red 1 x 20µl
    CB Reaction Buffer NM 1 x 5ml
    Substrate Ac-RR-AFC (10mM) Brown 1 x 200µl
  • 研究领域
  • 相关性Cathepsin B is a normal lysosomal proteinase that is expressed in all cells. It is a cysteine endoproteinase structurally and functionally related to the papain family of proteases. Cathepsin B is synthesized as an inactive pro enzyme which, by removal of a 62 amino acid propeptide, is activated to the single chain form of 31 kDa or the two chain form (25 and 5 kDa subunits). Matured Cathepsin B is normally localized in the lysosomes where it functions in protein turnover. Cathepsin B is also shown capable of degrading extracellular matrix proteins at acidic and neutral pH. Endogenous inhibitors of cysteine proteases, the cystatins and stefins, may play a major role in regulating Cathepsin B activity. In tumor cells and in cells exposed to mitogens, cathepsin B displays altered cellular trafficking resulting in the secretion of the 43 kDa precursor. Elevated levels of Cathepsin B correlate with malignancy suggesting that this enzyme may be a useful prognostic marker for several types of human cancers. Cathepsin B has also been implicated in the pathogenesis of rhematoid arthritis, muscular dystrophy and tumour metastasis. It is also known as amyloid precursor protein secretase and is involved in the proteolytic processing of amyloid precursor protein (APP).
  • 细胞定位Lysosome. Melanosome. Note=Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • 别名
    • Amyloid precursor protein secretase
    • APP secretase
    • APPS
    • Cathepsin B1
    • CathepsinB
    • CPSB
    • CTSB
    • Preprocathepsin B
    see all

应用

Our Abpromise guarantee covers the use of ab65300 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.

Cathepsin B Activity Assay Kit (Fluorometric) 图像

  • Cathepsin B activity in HL60 cell lysates (2.5x10e6 cells) with the addition of inhibitor. Background signal subtracted, duplicates; +/- SD.

实验方案

Cathepsin B Activity Assay Kit (Fluorometric) (ab65300)参考文献

This product has been referenced in:
  • Dautzenberg IJ  et al. Mammalian orthoreovirus T3D infects U-118 MG cell spheroids independent of junction adhesion molecule-A. Gene Ther 21:609-17 (2014). Functional Studies . Read more (PubMed: 24739522) »
  • Jang BG  et al. Impairment of Autophagic Flux Promotes Glucose Reperfusion-Induced Neuro2A Cell Death after Glucose Deprivation. PLoS One 8:e76466 (2013). Read more (PubMed: 24124562) »

See all 7 Publications for this product

Product Wall

Thank you for your enquiry. It may be possible to use plasma samples with this kit plasma. To date, however, we have no data testing plasma samples with this kit. If you do decide to use ab65300 with plasma samples I'd encourage you to submit an Abrevi...

Read More

I'm sorry we cannot share the composition of the lysis or the reaction buffer due to proprietary reasons. However, the lysis buffer contains Tween 20 which might not always work well with all enzyme-based assays.

Thank you for contacting us.

The sensitivity of these cathepsins is: 10 – 100 ng/assay.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.


The lab let me know that they do not have any additional tips for tissue processing other than the ones we mention on the FAQ site.


12. How should tissue be prepared for the kits?

Samples should be completely homogenized, pr...

Read More

Free AFC is available as product ID ab129872.

Thank you for your enquiry.

I can confirm this ab65300 Cathepsin B Activity Assay Kit (Fluorometric) is compatible with all mammalian samples, so it can be used with mice and rat samples. Here are a few citations which I hope may be benefici...

Read More

Thank you for contacting us.

I have had the chance to speak with the lab regarding your question. Each buffer is of different composition and they cannot be used interchangeably.

I hope this information is helpful to you. Plea...

Read More

Thank you for contacting us and your interest in our products.

The Cathepsin B Activity Fluorometric Assay Kit (ab65300) has not currently been tested with plasma samples.However, as long as theplasma samples contain Cathepsin B within the de...

Read More

According to our records, ab65300 was proving difficult to use and we were in contact in order to help resolve the issue. Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the diff...

Read More

Thanks for your call today and for letting us know about the problem with ab65300. As we discussed, please send your data and I will forward it on to the lab to see if there is any explanation or suggestion for your results. Could you also tell ...

Read More

1-10 of 12 Abreviews or Q&A

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