Cathepsin D Inhibitor Assay Kit (ab126779)
Key features and details
- Assay type: Quantitative
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 1 hr
- Sample type: Inhibitor compounds
Overview
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Product name
Cathepsin D Inhibitor Assay Kit
See all Cathepsin D kits -
Detection method
Fluorescent -
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).
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Notes
This product is manufactured by BioVision, an Abcam company and was previously called K148 Cathepsin D Inhibitor Screening Kit (Fluorometric). K148-100 is the same size as the 100 test size of ab126779.
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.
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Platform
Microplate reader
Properties
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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 Amber 1 x 0.2ml Pepstatin A (10 µM in DMSO) Blue 1 x 20µl -
Research areas
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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
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Related Products
Images
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Functional Studies - Cathepsin D Inhibitior Screening Kit (ab126779)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.
Datasheets and documents
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SDS download
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Datasheet download
References (1)
ab126779 has been referenced in 1 publication.
- Qiao S et al. The antimalarial amodiaquine causes autophagic-lysosomal and proliferative blockade sensitizing human melanoma cells to starvation- and chemotherapy-induced cell death. Autophagy 9:2087-102 (2013). PubMed: 24113242