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Cathepsin D Activity Assay Kit ab65302 is a fluorescence-based assay that uses the preferred cathepsin-D substrate sequence GKPILFFRLK(Dnp)-D-R-NH2) labeled with MCA.

Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.


Images

Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (AB65302), expandable thumbnail
  • Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (AB65302), expandable thumbnail
  • Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (AB65302), expandable thumbnail
  • Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (AB65302), expandable thumbnail
  • Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (AB65302), expandable thumbnail

Publications

Key facts

Detection method

Fluorescent

Sample types

Tissue Extracts, Cell Lysate

Assay type

Semi-quantitative

Reactive species

Mammals

Assay time

2h

Associated Products

Select an associated product type

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What's included?

2000 Test
Components
CD Cell Lysis Buffer
20 x 100 mL
CD Reaction Buffer
20 x 5 mL
Substrate II
20 x 200 µL

Recommended products

Cathepsin D Activity Assay Kit ab65302 is a fluorescence-based assay that uses the preferred cathepsin-D substrate sequence GKPILFFRLK(Dnp)-D-R-NH2) labeled with MCA.

Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.

Key facts

Detection method

Fluorescent

Sample types

Tissue Extracts, Cell Lysate

Assay type

Semi-quantitative

Reactive species

Mammals

Assay time

2h

Assay Platform

Microplate reader

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

Storage information

-20°C

Notes

Cathepsin D Activity Assay Kit ab65302 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.

This product is manufactured by BioVision, an Abcam company and was previously called K143 Cathepsin D Activity Fluorometric Assay Kit. K143-100 is the same size as the 100 test size of ab65302.

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Cathepsin D also known as CTSD is a protein with a mass of approximately 45 kDa. It functions as an aspartyl protease and is expressed in lysosomes across various tissues including the liver and kidneys. This enzyme acts by cleaving peptide bonds within proteins which is essential for protein degradation and turnover. Cathepsin D exists as precursor forms that become activated in the acidic environment of the lysosome. It plays a critical role in normal cellular processes by maintaining protein homeostasis.

Biological function summary

The enzymatic activity of Cathepsin D is important for cellular maintenance and apoptosis. This protease does not act within larger protein complexes but contributes to the degradation of extracellular and intracellular proteins. It mediates processes like antigen processing where it deconstructs proteins into peptides that are presented on major histocompatibility complex (MHC) molecules. ELISA tests can quantify its expression levels sometimes termed as CTSD activity in various biological samples offering insights into its role within cellular environments.

Pathways

Cathepsin D involvement includes the lysosomal degradation pathway and the apoptotic signaling pathway. In the lysosomal degradation pathway Cathepsin D breaks down proteins and peptides a process important for cellular recycling and energy release. It interacts with other lysosomal enzymes such as Cathepsin B in this pathway ensuring comprehensive breakdown of cellular waste. The apoptotic signaling pathway involves the regulation of programmed cell death where Cathepsin D can influence the activation of downstream proteins like Bcl-2 and Bax which control cell survival.

Associated diseases and disorders

The overexpression of Cathepsin D links to breast cancer and Alzheimer's disease. In breast cancer increased Cathepsin D expression correlates with tumor progression and metastasis influencing tumor behavior through interactions with other proteins involved in cell proliferation. Alzheimer's disease features the involvement of Cathepsin D in the breakdown of amyloid precursor protein which relates to amyloid beta plaque accumulation. The abnormal activity of Cathepsin D in these disorders makes it a potential target for therapeutic antibodies such as CTSD antibodies which aim to regulate its function.

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5 product images

  • Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (ab65302), expandable thumbnail
    Bong G.J., PLoS One 8(10), Fig 5F. doi:10.1371/journal.pone.0076466 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (ab65302)

    Enzyme activity of Cathepsin B and Cathepsin D was determined using Cathepsin B/D activity assay kit (Cathepsin B Activity Assay Kit (Fluorometric) ab65300 and ab65302). Neuro2a cell lysates were centrifuged at 10,000 xg for 10 minutes at 4°C 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).

  • Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (ab65302), expandable thumbnail

    Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (ab65302)

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

  • Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (ab65302), expandable thumbnail

    Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (ab65302)

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

  • Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (ab65302), expandable thumbnail

    Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (ab65302)

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

  • Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (ab65302), expandable thumbnail

    Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (ab65302)

    Cathepsin D measured in Jurkat cells.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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