Cathepsin B Activity Assay Kit (Fluorometric) (ab65300)
Key features and details
- Assay type: Quantitative
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 2 hr
- Sample type: Cell Lysate, Tissue Extracts
製品の概要
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製品名
Cathepsin B Activity Assay Kit (Fluorometric)
Cathepsin B キット 製品一覧 -
検出方法
Fluorescent -
サンプルの種類
Tissue Extracts, Cell Lysate -
アッセイタイプ
Quantitative -
全工程の試験時間
2h 00m -
製品の概要
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 37ºC
- analyze with microplate reader -
特記事項
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.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the SDS download section.
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試験プラットフォーム
Microplate reader
製品の特性
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保存方法
Store at -20°C. Please refer to protocols. -
内容 100 tests Cathepsin B Cell Lysis Buffer 1 x 25ml Cathepsin Inhibitor 1 x 20µl Cathepsin B Reaction Buffer 1 x 5ml Cathepsin B Substrate 1 x 200µl -
研究分野
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関連性
Cathepsin B is a normal lysosomal proteinase that is expressed in all cells. It is a cysteine endoproteinase structurally and functionally related to the papain family of proteases. Cathepsin B is synthesized as an inactive pro enzyme which, by removal of a 62 amino acid propeptide, is activated to the single chain form of 31 kDa or the two chain form (25 and 5 kDa subunits). Matured Cathepsin B is normally localized in the lysosomes where it functions in protein turnover. Cathepsin B is also shown capable of degrading extracellular matrix proteins at acidic and neutral pH. Endogenous inhibitors of cysteine proteases, the cystatins and stefins, may play a major role in regulating Cathepsin B activity. In tumor cells and in cells exposed to mitogens, cathepsin B displays altered cellular trafficking resulting in the secretion of the 43 kDa precursor. Elevated levels of Cathepsin B correlate with malignancy suggesting that this enzyme may be a useful prognostic marker for several types of human cancers. Cathepsin B has also been implicated in the pathogenesis of rhematoid arthritis, muscular dystrophy and tumour metastasis. It is also known as amyloid precursor protein secretase and is involved in the proteolytic processing of amyloid precursor protein (APP). -
細胞内局在
Lysosome. Melanosome. Note=Identified by mass spectrometry in melanosome fractions from stage I to stage IV. -
別名
- Amyloid precursor protein secretase
- APP secretase
- APPS
see all
関連製品
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Related Products
画像
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Functional studies - Cathespin B Activity Assay KitBong 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/
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 4ºC 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).
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Functional assays: Cathepsin B Activity Assay Kit (Fluorometric) (ab65300)Cathepsin B activity in HL60 cell lysates (2.5x10e6 cells) with the addition of inhibitor.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (98)
ab65300 は 98 報の論文で使用されています。
- Aly R et al. Dysregulation of kidney proteases in the pathogenesis of hypertension following unilateral nephrectomy in juvenile mice. Am J Transl Res 16:544-556 (2024). PubMed: 38463588
- Huang KC et al. Capsaicin alleviates cisplatin-induced muscle loss and atrophy in vitro and in vivo. J Cachexia Sarcopenia Muscle 14:182-197 (2023). PubMed: 36401337
- Natan Y et al. Amorphous Calcium Carbonate Shows Anti-Cancer Properties That are Attributed to Its Buffering Capacity. Cancers (Basel) 15:N/A (2023). PubMed: 37568601
- Wu Y et al. 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. Acta Neuropathol Commun 11:132 (2023). PubMed: 37580797
- Wang P et al. α-Synuclein-carrying astrocytic extracellular vesicles in Parkinson pathogenesis and diagnosis. Transl Neurodegener 12:40 (2023). PubMed: 37620916