Cathepsin B Activity Assay kit (Fluorometric) (ab65300) is a fluorescence-based assay that uses the preferred cathepsin-B substrate sequence RR labeled with AFC (amino-4-trifluoromethyl coumarin).
Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
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- American journal of translational research 16:544-5562024Dysregulation of kidney proteases in the pathogenesis of hypertension following unilateral nephrectomy in juvenile mice.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRasha Aly,Yunus E Dogan,Niharika Bala,Carlos Lugo,Seena Darwish,Lawrence Shoemaker,Abdel A AlliPubMed 38463588
- Cell communication and signaling : CCS 21:2752023The Golgi stacking protein GRASP55 is targeted by the natural compound prodigiosin.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLena Berning,Thomas Lenz,Ann Kathrin Bergmann,Gereon Poschmann,Hannah U C Brass,David Schlütermann,Annabelle Friedrich,María José Mendiburo,Céline David,Seda Akgün,Jörg Pietruszka,Kai Stühler,Björn StorkPubMed 37798768
- Translational neurodegeneration 12:402023α-Synuclein-carrying astrocytic extracellular vesicles in Parkinson pathogenesis and diagnosis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesPan Wang,Guoyu Lan,Bin Xu,Zhenwei Yu,Chen Tian,Xia Lei,Wassilios G Meissner,Tao Feng,Ying Yang,Jing ZhangPubMed 37620916
- Acta neuropathologica communications 11:1322023Cathepsin B abundance, activity and microglial localisation in Alzheimer's disease-Down syndrome and early onset Alzheimer's disease; the role of elevated cystatin B.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYixing Wu,Paige Mumford,Suzanna Noy,Karen Cleverley,Alicja Mrzyglod,Dinghao Luo,Floris van Dalen,Martijn Verdoes,Elizabeth M C Fisher,Frances K WisemanPubMed 37580797
- Cancers 15:2023Amorphous Calcium Carbonate Shows Anti-Cancer Properties That are Attributed to Its Buffering Capacity.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYehudit Natan,Yigal Dov Blum,Amir Arav,Ylena Poliansky,Sara Neuman,Orit Ecker Cohen,Yossi BenPubMed 37568601
- Disease models & mechanisms 16:2023Oxidative stress induces lysosomal membrane permeabilization and ceramide accumulation in retinal pigment epithelial cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKevin 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 DunaiefPubMed 37401371
- Pharmaceuticals (Basel, Switzerland) 15:2022Toosendanin Induces Hepatocyte Damage by Inhibiting Autophagic Flux via TFEB-Mediated Lysosomal Dysfunction.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLi Luo,Yonghong Liang,Yuanyuan Fu,Zhiyuan Liang,Jinfen Zheng,Jie Lan,Feihai Shen,Zhiying HuangPubMed 36558960
- Journal of cachexia, sarcopenia and muscle 14:182-1972022Capsaicin alleviates cisplatin-induced muscle loss and atrophy in vitro and in vivo.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKo-Chieh Huang,Yi-Fen Chiang,Tsui-Chin Huang,Hsin-Yuan Chen,Po-Han Lin,Mohamed Ali,Shih-Min HsiaPubMed 36401337
- NPJ Parkinson's disease 8:1262022Lysosomal lipid alterations caused by glucocerebrosidase deficiency promote lysosomal dysfunction, chaperone-mediated-autophagy deficiency, and alpha-synuclein pathology.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAlba Navarro-Romero,Irene Fernandez-Gonzalez,Jordi Riera,Marta Montpeyo,Merce Albert-Bayo,Tresa Lopez-Royo,Pablo Castillo-Sanchez,Clara Carnicer-Caceres,Jose Antonio Arranz-Amo,Laura Castillo-Ribelles,Eddie Pradas,Josefina Casas,Miquel Vila,Marta Martinez-VicentePubMed 36202848
- Signal transduction and targeted therapy 7:3172022Elaiophylin triggers paraptosis and preferentially kills ovarian cancer drug-resistant cells by inducing MAPK hyperactivation.Applications:Unspecified applicationReactive species:Unspecified reactive speciesGuan-Nan Li,Xue-Jiao Zhao,Zhen Wang,Meng-Shi Luo,Shen-Nan Shi,Dan-Mei Yan,Hua-Yi Li,Jia-Hao Liu,Yang Yang,Jia-Hong Tan,Ze-Yu Zhang,Ru-Qi Chen,Hui-Ling Lai,Xiao-Yuan Huang,Jian-Feng Zhou,Ding Ma,Yong Fang,Qing-Lei GaoPubMed 36097006
Key facts
- Detection method
Fluorescent
- Sample types
Tissue Extracts, Cell Lysate
- Assay type
Quantitative
- Assay time
2h
Associated Products
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Target data
Function
Thiol protease which is believed to participate in intracellular degradation and turnover of proteins (PubMed:12220505). Cleaves matrix extracellular phosphoglycoprotein MEPE (PubMed:12220505). Involved in the solubilization of cross-linked TG/thyroglobulin in the thyroid follicle lumen (By similarity). Has also been implicated in tumor invasion and metastasis (PubMed:3972105).
Alternative names
Cathepsin B, APP secretase, Cathepsin B1, APPS, CTSB, CPSB
What's included?
Recommended products
Cathepsin B Activity Assay kit (Fluorometric) (ab65300) is a fluorescence-based assay that uses the preferred cathepsin-B substrate sequence RR labeled with AFC (amino-4-trifluoromethyl coumarin).
Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
Alternative names
Cathepsin B, APP secretase, Cathepsin B1, APPS, CTSB, CPSB
Key facts
- Detection method
Fluorescent
- Sample types
Tissue Extracts, Cell Lysate
- Assay type
Quantitative
- 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 B Activity Assay kit (Fluorometric) (ab65300) is a fluorescence-based assay that utilizes the preferred cathepsin-B substrate sequence RR labeled with AFC (amino-4-trifluoromethyl coumarin).
Cell lysates or other samples that contain cathepsin-B will cleave the synthetic substrate RR-AFC to release free AFC. The released AFC can easily be quantified using a fluorometer or fluorescence plate reader at Ex/Em 400/505 nm.
The Cathepsin B assay protocol is simple, straightforward, and can be adapted to 96-well plate assays. Assay conditions have been optimized to obtain the maximal activity.
Cathepsin B assay protocol summary:
- add samples to wells
- add reaction mix and incubate for 1-2 hr at 37°C
- analyze with microplate reader
This product is manufactured by BioVision, an Abcam company and was previously called K140 Cathepsin B Activity Fluorometric Assay Kit. K140-100 is the same size as the 100 test size of ab65300.
Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Cathepsin B also known as CTSB or cathepsin beta is a lysosomal cysteine protease belonging to the papain family. This enzyme possesses a molecular weight of approximately 25 to 30 kDa. You can find cathepsin B expressed in various tissues with high levels observed in the liver kidney and spleen. It plays a mechanical role in protein degradation within lysosomes operating optimally in acidic environments. The enzyme has active and inactive forms depending on structural modifications and environmental conditions.
- Biological function summary
Cathepsin B participates as a proteolytic enzyme involved in protein turnover and processing. It does not function in isolation but often associates with various other proteases forming complexes essential for efficient substrate breakdown. Cathepsin B assists in the activation of other enzymes by cleaving propeptides therefore facilitating the initiation of cascade reactions. This role is critical in maintaining cellular homeostasis and modulating physiological processes including cell migration and immune response modulation.
- Pathways
Cathepsin B significantly contributes to the lysosomal protein degradation pathway and the autophagy-lysosome pathway. It interacts with proteins such as B protein which are important for cellular recycling and energy homeostasis. Its activity supports the MHC class II antigen processing pathway playing an essential part in immune system functionality. Cathepsin B also exhibits interactions with Ctsb protein inhibitors which regulate its proteolytic activity within these pathways.
- Associated diseases and disorders
Cathepsin B has strong associations with cancer progression and neurological disorders such as Alzheimer's disease. Its overexpression and dysregulation are linked to tumor metastasis where it degrades extracellular matrix components promoting invasion. Ctsb protein interactions with corey mazo proteins influence tumor microenvironment and immune evasion. In Alzheimer's disease cathepsin B can influence amyloid beta peptide processing connected to neurodegenerative pathology progression. These roles highlight the potential for cathepsin B as a therapeutic target in addressing these conditions.
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2 product images
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 B Activity Assay Kit (Fluorometric) (ab65300)
Functional studies - Cathespin B Activity Assay Kit.
Enzyme activity of Cathepsin B and Cathepsin D was determined using Cathepsin B/D activity assay kit (ab65300 and Cathepsin D Activity Assay Kit (Fluorometric) 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 B Activity Assay Kit (Fluorometric) (ab65300)
Cathepsin B activity in HL60 cell lysates (2.5x10e6 cells) with the addition of inhibitor.
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