Overview

  • Product name
    Cathepsin D Inhibitor Screening Kit
    See all Cathepsin D kits
  • Sample type
    Inhibitor compounds
  • Assay type
    Quantitative
  • Assay time
    1h 00m
  • Product overview

    Cathepsin D Inhibitor Screening Kit (ab126779) is a fluorescence-based assay that utilizes the preferred cathepsin D substrate sequence GKPILFFRLK(Dnp)-D-R-NH2) labeled with MCA. Cathepsin D will cleave the synthetic substrate to release the quenched fluorescent group MCA, which can then easily be measured using a fluorometer or fluorescence plate reader at Ex/Em = 328/460 nm. The relative efficacy of test inhibitors are compared to the positive control inhibitor, Pepstatin A (IC50 < 0.1 nM). The Cathepsin D assay is simple, straightforward, and can be adapted to 96-well plate assays and is suitable for high throughput screening (HTS).
    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 pro-apoptotic signals. Cathepsins are lysosomal enzymes that are also used as sensitive markers in various toxicological investigations.

  • Tested applications
    Suitable for: Functional Studiesmore details

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components Identifier 100 tests
    Cathepsin D, human (Lyophilized) Green 1 vial
    CD Reaction Buffer NM 1 x 10ml
    CD Substrate Brown 1 x 0.2ml
    Pepstatin A (10 µM in DMSO) Blue 1 x 20µl
  • Research areas
  • Relevance
    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.
  • Cellular localization
    Lysosome. Melanosome. Note=Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Alternative names
    • CLN10
    • CPSD
    • CTSD
    • Lysosomal aspartyl protease

Associated products

Applications

Our Abpromise guarantee covers the use of ab126779 in the following tested applications.

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

Application Abreviews Notes
Functional Studies Use at an assay dependent dilution.

Images

  • Typical Pepstatin A Inhibition Profile of Cathepsin D Activity. The red line denotes an IC50 value of 0.067 nM. Results were analyzed by fluorescence plate reader according to the kit instructions.

Protocols

References

ab126779 has not yet been referenced specifically in any publications.

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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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