Caspase-3 Assay Kit (Fluorometric) (ab39383)
Key features and details
- Assay type: Enzyme activity
- 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
Caspase-3 Assay Kit (Fluorometric)
Caspase-3 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
Enzyme activity -
全工程の試験時間
2h 00m -
製品の概要
Caspase-3 Assay Kit (Fluorometric) (ab39383) provides a simple and convenient means for assaying DEVD-dependent caspase activity.
The Caspase-3 assay protocol is based on detection of cleavage of substrate DEVD-AFC (AFC: 7-amino-4-trifluoromethyl coumarin).
DEVD-AFC emits blue light (λ max = 400 nm). On cleavage of the substrate by CPP32 or related caspases, free AFC emits a yellow-green fluorescence (Ex/Em = 400/505 nm).
The signal can be quantified using a fluorometer or a fluorescence microtiter plate reader. Comparison of the fluorescence of AFC from an apoptotic sample with an uninduced control allows determination of the fold increase in Caspase-3/CPP32 activity.
Caspase-3 assay protocol summary:
- lyse cells / homogenize and lyse tissues in lysis buffer
- incubate on ice for 10 min
- add reaction buffer and DEVD-AFC substrate and incubate for 1-2 hr at 37ºC
- analyze with fluorometer or microplate reader -
特記事項
This product is manufactured by BioVision, an Abcam company and was previously called K105 Caspase-3 Fluorometric Assay Kit. K105-100 is the same size as the 100 test size of ab39383.
Due to the nature of the substrate, this assay also detects caspase-7 activity.
Other caspase and apoptosis assays
Review the full set of caspase assays, or the apoptosis assay and apoptosis marker guide.
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.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 2X Reaction Buffer I 4 x 2ml DEVD-AFC 1 x 500µl DTT I 1 x 400µl Lysis Buffer IV 1 x 100ml -
研究分野
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機能
Involved in the activation cascade of caspases responsible for apoptosis execution. At the onset of apoptosis it proteolytically cleaves poly(ADP-ribose) polymerase (PARP) at a '216-Asp-
-Gly-217' bond. Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain. Cleaves and activates caspase-6, -7 and -9. Involved in the cleavage of huntingtin. -
組織特異性
Highly expressed in lung, spleen, heart, liver and kidney. Moderate levels in brain and skeletal muscle, and low in testis. Also found in many cell lines, highest expression in cells of the immune system. -
配列類似性
Belongs to the peptidase C14A family. -
翻訳後修飾
Cleavage by granzyme B, caspase-6, caspase-8 and caspase-10 generates the two active subunits. Additional processing of the propeptides is likely due to the autocatalytic activity of the activated protease. Active heterodimers between the small subunit of caspase-7 protease and the large subunit of caspase-3 also occur and vice versa.
S-nitrosylated on its catalytic site cysteine in unstimulated human cell lines and denitrosylated upon activation of the Fas apoptotic pathway, associated with an increase in intracellular caspase activity. Fas therefore activates caspase-3 not only by inducing the cleavage of the caspase zymogen to its active subunits, but also by stimulating the denitrosylation of its active site thiol. -
細胞内局在
Cytoplasm. - Information by UniProt
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別名
- A830040C14Rik
- Apopain
- CASP 3
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関連製品
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Related Products
画像
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Functional assays: Caspase-3 assay kit (ab39383)Image from Lin S.F et al., PLoS One 12(2). Fig4B. doi: 10.1371/journal.pone.0172315. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/
Caspase-3 activity was analyzed using fluorometric assay kit (ab39383). Thyroid cancer cells were plated at 1 x 106 cells in 10 mL of media and incubated overnight. Cells were treated with Dinaciclib (25 nM) for 24 hours. Adherent cells (5 x 105) were collected, centrifuged, lysed using 50 μL of lysis buffer on ice for 10 min, incubated with DEVD-AFC substrate and reaction buffer at 37°C for 1.5 hours. Caspase-3 activity was detected by spectrophotometry. The fluorescence intensity of the treated samples was compared with that of control samples to determine the fold-increase in caspase activity. Each condition was performed in duplicate.
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Functional assays: Caspase-3 Assay Kit (Fluorometric) (ab39383)Caspase-3 activity in Jurkat lysates (6.6 x10e5 cells) following 20 hour exposure to 2 uM Camptothecin (ab120115) or 10 ng/mL anti-Fas Ab (MBL). Background signal subtracted, duplicates +/- SD.
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 (96)
ab39383 は 96 報の論文で使用されています。
- Alharthy RD et al. Pyrazole derivatives of pyridine and naphthyridine as proapoptotic agents in cervical and breast cancer cells. Sci Rep 13:5370 (2023). PubMed: 37005457
- Frost J et al. Telaprevir Treatment Reduces Paralysis in a Mouse Model of Enterovirus D68 Acute Flaccid Myelitis. J Virol 97:e0015623 (2023). PubMed: 37154751
- Marshall SK et al. Evaluation of Single Dose and Fractionated Dose of I-131 Radiolabeled Nanoparticles for Triple-Negative Breast Cancer Treatment. Biomedicines 11:N/A (2023). PubMed: 37626666
- Grafals-Ruiz N et al. MicroRNA-92b targets tumor suppressor gene FBXW7 in glioblastoma. Front Oncol 13:1249649 (2023). PubMed: 37752997
- Beillevaire D et al. Autolysosomes and caspase-3 control the biogenesis and release of immunogenic apoptotic exosomes. Cell Death Dis 13:145 (2022). PubMed: 35149669