AB65302

Cathepsin D Activity Assay Kit (Fluorometric)

Name: Cathepsin D Activity Assay Kit (Fluorometric) (ab65302/K143-100) | アブカム
Brand: Abcam Limited
SKU: AB65302
Rating: 4 (3 reviews)

(3 Reviews)

(127 Publications)

Cathepsin D Activity Assay Kit (Fluorometric) ab65302 uses a substrate based on the GKPILFFRLK(Dnp)-D-R-NH2) Cathepsin D substrate. Add reaction mix to samples and incubate for 1-2 hr at 37°C. Readout on any fluorometric (Ex/Em 328/460 nm) plate reader.

- Cited in > 120 publications
- Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.

5 Images

Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (AB65302)

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Lab

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).

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Supplier Data

Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (AB65302)

Cathepsin D measured in Jurkat cells.

FuncS

Lab

Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (AB65302)

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

FuncS

Lab

Functional Studies - Cathepsin D Activity Assay Kit (Fluorometric) (AB65302)

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

FuncS

PubMed

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 (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).

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/

Key facts

検出方法

Fluorescent

サンプルタイプ

Tissue Extracts, Cell Lysate

アッセイタイプ

Semi-quantitative

アッセイ時間

アッセイプラットフォーム

Microplate reader

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "Enzyme activity assay": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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製品の詳細

Cathepsin D Activity Assay Kit (Fluorometric) ab65302 is a fluorescence-based assay to measure Cathepsin D.

How the assay works

Cathepsin D Activity Assay Kit (Fluorometric) 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.

Cathepsin D assay protocol summary:

-Add samples to wells
- Add reaction mix and incubate for 1-2 hr at 37°C
- Measure fluorescence (Ex/Em = 328/460 nm)

Related and recommended products

See below other alternative Cathepsin Assay Kits:
- Cathepsin L Assay Kit (Magic Red) ab270774
- Cathepsin K Assay Kit (Magic Red) ab270774

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.

The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

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製品内容

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出荷温度及び保存条件

出荷温度

Blue Ice

短期保存温度

-20°C

長期保存温度

-20°C

保管に関する情報

-20°C

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補足情報

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

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.

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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製品プロトコール

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ターゲットの情報

See full target information Cathepsin D

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文献 (127)

Recent publications for all applications. Explore the full list and refine your search

American journal of translational research 16:544-556 PubMed38463588

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Dysregulation of kidney proteases in the pathogenesis of hypertension following unilateral nephrectomy in juvenile mice.

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Rasha Aly,Yunus E Dogan,Niharika Bala,Carlos Lugo,Seena Darwish,Lawrence Shoemaker,Abdel A Alli

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Inhalation of acidic nanoparticles prevents doxorubicin cardiotoxicity through improvement of lysosomal function.

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Yohan Santin,Karina Formoso,Fraha Haidar,Maria Del Pilar Oreja Fuentes,Florence Bourgailh,Nesrine Hifdi,Karim Hnia,Yosra Doghri,Jessica Resta,Camille Champigny,Séverine Lechevallier,Maximin Détrait,Grégoire Cousin,Malik Bisserier,Angelo Parini,Frank Lezoualc'h,Marc Verelst,Jeanne Mialet-Perez

Translational neurodegeneration 12:40 PubMed37620916

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α-Synuclein-carrying astrocytic extracellular vesicles in Parkinson pathogenesis and diagnosis.

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Pan Wang,Guoyu Lan,Bin Xu,Zhenwei Yu,Chen Tian,Xia Lei,Wassilios G Meissner,Tao Feng,Ying Yang,Jing Zhang

Acta neuropathologica communications 11:132 PubMed37580797

2023

Cathepsin B abundance, activity and microglial localisation in Alzheimer's disease-Down syndrome and early onset Alzheimer's disease; the role of elevated cystatin B.

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Yixing Wu,Paige Mumford,Suzanna Noy,Karen Cleverley,Alicja Mrzyglod,Dinghao Luo,Floris van Dalen,Martijn Verdoes,Elizabeth M C Fisher,Frances K Wiseman

Disease models & mechanisms 16: PubMed37401371

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Oxidative stress induces lysosomal membrane permeabilization and ceramide accumulation in retinal pigment epithelial cells.

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Kevin R Zhang,Connor S R Jankowski,Rayna Marshall,Rohini Nair,Néstor Más Gómez,Ahab Alnemri,Yingrui Liu,Elizabeth Erler,Julia Ferrante,Ying Song,Brent A Bell,Bailey H Baumann,Jacob Sterling,Brandon Anderson,Sierra Foshe,Jennifer Roof,Hossein Fazelinia,Lynn A Spruce,Jen-Zen Chuang,Ching-Hwa Sung,Anuradha Dhingra,Kathleen Boesze-Battaglia,Venkata R M Chavali,Joshua D Rabinowitz,Claire H Mitchell,Joshua L Dunaief

European journal of oral sciences 131:e12923 PubMed36788303

2023

Role of cathepsin D induced by Porphyromonas gingivalis lipopolysaccharide in periodontitis.

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Hyun Woong Jeong,Dong Sik Chang,June Soo Kim,Young Sun Hwang

International journal of molecular sciences 24: PubMed36768815

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"Pulsed Hypoxia" Gradually Reprograms Breast Cancer Fibroblasts into Pro-Tumorigenic Cells via Mesenchymal-Epithelial Transition.

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Anna Nushtaeva,Mikhail Ermakov,Maria Abdurakhmanova,Olga Troitskaya,Tatyana Belovezhets,Mikhail Varlamov,Tatyana Gayner,Vladimir Richter,Olga Koval

Cells 12: PubMed36672225

2023

Chloroquine Enhances Death in Lung Adenocarcinoma A549 Cells Exposed to Cold Atmospheric Plasma Jet.

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Ekaterina Patrakova,Mikhail Biryukov,Olga Troitskaya,Pavel Gugin,Elena Milakhina,Dmitriy Semenov,Julia Poletaeva,Elena Ryabchikova,Diana Novak,Nadezhda Kryachkova,Alina Polyakova,Maria Zhilnikova,Dmitriy Zakrevsky,Irina Schweigert,Olga Koval

Cell communication and signaling : CCS 20:192 PubMed36474295

2022

Catalase-deficient mice induce aging faster through lysosomal dysfunction.

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Raghbendra Kumar Dutta,Joon No Lee,Yunash Maharjan,Channy Park,Seong-Kyu Choe,Ye-Shih Ho,Hyug Moo Kwon,Raekil Park

International journal of molecular sciences 23: PubMed36233303

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Saposin C, Key Regulator in the Alpha-Synuclein Degradation Mediated by Lysosome.

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Clara Ruz,Francisco J Barrero,Javier Pelegrina,Sara Bandrés-Ciga,Francisco Vives,Raquel Duran

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Cathepsin D Activity Assay Kit (Fluorometric)