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#5723 Caspase-3 Activity Assay Kit

  • Caspase-3 Activity Assay Kit Data Sheet PDF
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希望納入価格 (円)
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2019年8月20日15時55分 現在
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#5723S1 Kit110,000
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CASPASE-3 製品一覧

キット内容 容量
Ac-DEVD-AMC 1 x 1 mg
AMC (7-amino-4-methylcoumarin) 1 x 250 µl
PathScan® Sandwich ELISA Lysis Buffer (1X) #7018 1 x 30 ml
Caspase Assay Buffer (2x) 1 x 30 ml
DTT (Dithiothreitol) #7016 1 x 192.8 mg
Kit 情報
細胞抽出物中のCaspase-3 の活性を検出する蛍光分析キットです。Caspase-3 の検出に必要な蛍光基質 (N-Acetyl-Asp-Glu-Val-Asp-7-amino-4-methylcoumarin またはAc-DEVD-AMC) を含みます。活性型Caspase-3 がこの基質をDEVD とAMC の間で切断し、高い蛍光性を持つAMC が生成され、これを蛍光リーダーで読み取ることでCaspase-3 の活性を測定します。
種交差性
すべての種
貯法
-20℃
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社内データ

Fluorescent Detection

Fluorescent Detection

Figure 1. NIH/3T3 cells were treated with Staurosporine #9953 (5 μM, 5 hr) and then lysed in PathScan® Sandwich ELISA Lysis Buffer (1X) #7018 (supplied with kit). Various amounts of cell lysate were added to assay plates containing the substrate solution, and plates were incubated at 37ºC in the dark. Relative fluorescent units (RFUs) were acquired at 1 and 4 hr.

Fluorescent Detection

Fluorescent Detection

Figure 2. NIH/3T3 cells were seeded in a 96-well plate at 1x105 cells/well or 5x104 cells/well, and then treated with Staurosporine #9953 (5 μM, 5 hr) and then lysed in 30 μl PathScan® Sandwich ELISA Lysis Buffer (1X) #7018 (supplied with kit). Cell lysate was added to assay plates containing the substrate solution, and plates were incubated at 37ºC in the dark. Relative fluorescent units (RFUs) were acquired at 0, 1, 2, 4, and 6 hr.

Fluorescent Detection

Fluorescent Detection

Figure 3. HeLa cells were seeded at 1x105 cells/well in a 96-well plate and incubated overnight. Cells were treated with various concentrations of Staurosporine #9953 (5 hr) and then lysed in 30 μL of PathScan® Sandwich ELISA Lysis Buffer (1X) #7018 (supplied with kit). Cell lysate was mixed with substrate solution and incubated at 37ºC in the dark for 2 hr and relative fluorescent units (RFUs) were acquired.


バックグラウンド

Caspase-3 (CPP-32, Apoptain, Yama, SCA-1) is a critical executioner of apoptosis, as it is either partially or totally responsible for the proteolytic cleavage of many key proteins, such as the nuclear enzyme poly (ADP-ribose) polymerase (PARP) (1). Activation of caspase-3 requires proteolytic processing of its inactive zymogen into activated p17 and p12 fragments. Cleavage of caspase-3 requires the aspartic acid residue at the P1 position (2).

Caspase-7 (CMH-1, Mch3, ICE-LAP3) has been identified as a major contributor to the execution of apoptosis (3-6). Caspase-7, like caspase-3, is an effector caspase that is responsible for cleaving downstream substrates, such as PARP (3,5). During apoptosis, caspase-7 is activated by upstream caspases through proteolytic processsing at Asp23, Asp198, and Asp206, thereby producing the mature subunits (3,5). Similar to caspases-2 and -3, caspase-7 preferentially cleaves substrates following the recognition sequence DEVD (7).

使用文献

PathScan is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

本製品は試験研究用です。

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