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Overview

  • Product nameCathepsin D Activity Assay Kit (Fluorometric)See all Cathepsin D kits ...
  • Tests
    100 x 1 assay
  • Sample type
    Tissue Extracts, Cell Lysate
  • Assay typeQuantitative
  • Assay time
    2h 00m
  • Product overview

    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.

  • Notes

    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.

  • Tested applicationsFunctional Studies more details

Properties

  • RelevanceCathepsin 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.
  • Cellular localizationLysosome. Melanosome. Note=Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
    • Alternative names
        CLN10CPSDCTSDLysosomal aspartyl protease

    Applications

    Our Abpromise guarantee covers the use of ab65302 in the following tested applications.

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

    Application Notes
    Functional Studies FuncS

    Cathepsin D Activity Assay Kit (Fluorometric) images

    • Functional Studies - Cathepsin D Activity Fluorometric Assay Kit (ab65302)
      Functional Studies - Cathepsin D Activity Fluorometric Assay Kit (ab65302)
      Cathepsin D assays were performed using various numbers of Jurkat Cells as indicated. Results were analyzed by fluorescence plate reader according to the kit instructions.

    Protocols

    References for Cathepsin D Activity Assay Kit (Fluorometric) (ab65302)

    This product has been referenced in:
    • Moles A  et al. Cathepsin B overexpression due to Acid sphingomyelinase ablation promotes liver fibrosis in niemann-pick disease. J Biol Chem 287:1178-88 (2012). Read more (PubMed: 22102288) »
    • Dehay B  et al. Loss of P-type ATPase ATP13A2/PARK9 function induces general lysosomal deficiency and leads to Parkinson disease neurodegeneration. Proc Natl Acad Sci U S A 109:9611-6 (2012). Read more (PubMed: 22647602) »

    See all 4 Publications for this product

    Publishing research using ab65302 ? Please let us know so that we can cite the reference in this datasheet

    Product Wall

    Submit an Abreview for ab65302 Submit a Question for ab65302

    Displaying 1 - 10 of 12 results for Abreviews and Q&A

    10
    Votes: Highest First

    HI

    We are looking at kits to measure Cathepsin B, D, and G in secretions (in PBS) from female reproductive tract. How best to adapt your kit to cell free samples?

    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 re...

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    Can we use 335 excitation filter instead of recomended 328?

    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)

    http://www.sciencedirect.com/science/article/pii/S01437208990...

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    Is the Substrate from this kit available separately?

    The catalog number for the substrate is ab154147 http://www.abcam.com/substrate-reagent-ab154147.html.

    Dear All,
    a customer of ours has bought your Cathepsin D Activity Assay Kit (Fluorometric) (ab65302).
    She would like to have confirmationthat it is possible to evaluate the activity of cathepsin D (in our case in samples of lymphomonocytes cell)...

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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 bu...

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    I am interested in measuring calpain and cathepsin activity in fish
    muscle. Are these kits suitable for that purpose?
    http://www.abcam.com/Calpain-Activity-Assay-Kit-ab65308.html
    http://www.abcam.com/Cathepsin-L-Activity-Assay-Kit-Fluo...

    Read More

    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 andy...

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    I order the cathepsin B and cathepsin D assay kits from Abcam. In the kit there are a cell lysis buffer without no identification about the content. I wanna use the same samples to measure both activities. Could I choose one cell lysis buffer or should I ...

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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. Please ...

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    Dear All, a customer of ours is interested in your ab65302 but she would like to know more info about the precision and the reliability of this kit.   Thanks in advance.

    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 quantified...

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    Thank you for your response. Can you send me some data corroborating your response? Thanks so much. best,

    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 D ...

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    Thank you for your response. Can you send me some data corroborating your response? Thanks so much. best,

    Thank you for your reply. I have asked the lab for any data or references that they might have regarding the specificity of the substrates, and I will forward this information to you once I hear from them. Please let me know if you have any ...

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    Do the cathepsin D and K activity assay kits cross-react with other enzymes?

    Thank you for your call yesterday and for your question about ab65302 and ab65303. I have heard back from the lab and we do not expect these kits to cross-react with other enzymes. I hope this information is useful, but please let me know if...

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