Cathepsin D Activity Assay Kit (Fluorometric) (ab65302)


  • 製品名
    Cathepsin D Activity Assay Kit (Fluorometric)
    Cathepsin D キット 製品一覧
  • サンプルの種類
    Tissue Extracts, Cell Lysate
  • アッセイタイプ
  • 全工程の試験時間
    2h 00m
  • 種交差性
    交差種: Mammals, Other species
  • 製品の概要

    Abcam's Cathepsin D Activity Assay Kit is a fluorescence-based assay that utilizes the preferred cathepsin-D substrate sequence GKPILFFRLK(Dnp)-D-R-NH2) labeled with MCA. Cell lysates or other samples that contain cathepsin-D will cleave the synthetic substrate to release fluorescence, which can then easily be quantified using a fluorometer or fluorescence plate reader at Ex/Em = 328/460 nm.
    Visit our FAQs page for tips and troubleshooting.

  • 特記事項

    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.


  • 保存方法
    Store at -20°C. Please refer to protocols.
  • 内容 ラベル 100 tests
    CD Cell Lysis Buffer WM 1 x 25ml
    CD Reaction Buffer NM 1 x 5ml
    CD Substrate (1mM) Brown 1 x 200µl
  • 研究分野
  • 関連性
    Cathepsin D is a normal lysosomal protease that is expressed in all cells. It is an aspartyl protease with a pH optimum in the range of 3-5, and contains two N linked oligosaccharides. Cathepsin D is synthesized as an inactive pro enzyme. Activation involves the proteolytic removal of the 43 amino acid profragment and an internal cleavage to generate the two chain form made up of 34 and 14 kDa subunits. Cathepsin D contains the mannose-6-phosphate lysosomal localization signal that targets the enzyme to the lysosomal compartment where it functions in the normal degradation of proteins. In certain tumor cells, Cathepsin D is abnormally processed and is secreted in its precursor form. Numerous clinical studies as well as in vitro evidence suggest that cathepsin D plays an important role in malignant transformation and may be a useful prognostic indicator for breast cancer and possibly Alzheimer's disease.
  • 細胞内局在
    Lysosome. Melanosome. Note=Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • 別名
    • CLN10
    • CPSD
    • CTSD
    • Lysosomal aspartyl protease



  • Enzyme activity of Cathepsin B and Cathepsin D was determined using Cathepsin B/D activity assay kit (ab65300 and ab65302). Neuro2a cell lysates were centrifuged at 10,000 xg for 10 minutes at 4ºC and supernatant was used for the assay. Cathepsin activity was performed at the 24 and 48 hours time points by cleavage of the fluorescence peptide substrate [DnP-DR-MCA, GKPILFFRLK(DnP)-DR substrate peptide labeled with MCA]. Data represent the mean ± SD (*p<0.05, **p<0.001).

  • Cathepsin D levels were measured in standard control samples from ab119586; background signal subtracted (duplicates +/- SD).

  • Cathepsin D measured in mouse tissue lysates (mg of extracted protein), background signal subtracted (duplicates +/- SD).

  • Cathepsin D measured in cell lysates, background signal subtracted (duplicates +/- SD).



This product has been referenced in:
  • Fu Z  et al. CQ sensitizes human pancreatic cancer cells to gemcitabine through the lysosomal apoptotic pathway via reactive oxygen species. Mol Oncol 12:529-544 (2018). Functional Studies ; Human . Read more (PubMed: 29453806) »
  • Tan K  et al. Expression and Localization of Cathepsins B, D, and G in Dupuytren's Disease. Plast Reconstr Surg Glob Open 6:e1686 (2018). Read more (PubMed: 29616179) »

See all 20 Publications for this product

レビューと Q&A

We do not recommend to prepare one lysate to use with ab65302, ab65306 and ab65307.
It is important to prepare separate lysates for each kit, using the specific lysis buffer provided. This is because the cathepsins have very specific pH and other c...

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Es preferible utilizar un pellet celular fresco, en lugar de uno congelado. De cualquier manera, si el pellet se ha almacenado a -80C inmediatamente después de lisar las células en principio no hay problema en utilizarlo con este kit. Hay...

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1) We have not tested adding increasing concentrations of DMSO with this kit.

I would definitely recommend adding protease inhibitor cocktail instead of DMSO, such as ab65621 Read More

Thank you for contacting us.

We have used this assay only with cell and tissue lysates, which will be lysed in the lysis buffer. To use with body fluid, I would recommend diluting it with the lysis buffer. The PBS should not interfere with the...

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Thank you for contacting us.

The fluorescent compound in this kit is MCA which has absorbance maxima ˜350 and excitation maxima ˜460. Please click the link for details (Fig 1)

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Thank you for your inquiry.

I can confirm that an inhibitor would be a good negative control for your customer. But it is also not necessary to to have a negative control for every sample.

This kit is not measuring absolute activity...

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Thank you for contacting Abcam.

We have not tested any of the kits with fish samples but theoretically as long as there are active enzymes in the sample, the kit should detect it. Fish cell lysates can be prepared as for the mammalian cells a...

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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...

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Thank you for your email and interest in Abcam products. The Cathepsin D Activity Assay Kit ab65302 is a simple and straightforward assay which is very sensitive as it is based on the detection of a fluorescent substrate which can easily be quantif...

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Thank you again for your question and for your patience. The lab has not specifically tested these kits against the other cathepsins, but the substrates have been thoroughly studied and are known to be the preferred substrates for cathepsin...

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