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Succinate-Glo™ JmjC Demethylase/Hydroxylase Assay

Part Numbers: V7990, V7991

V7990_Succinate-Glo-JmjC DemethylaseHydroxylase Assay
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高感度に脱メチル化酵素活性を発光で検出

  • 添加 - 測定だけの簡便なアッセイフォーマット
  • 実質的にコハク酸を生成するあらゆる酵素を測定できるユニバーサルアッセイ
  • スクリーニング用途に最適化され、偽陽性も最低限

Size

選択製品のカタログ番号: V7990

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Succinate-Glo™ JmjC Demethylase/Hydroxylase Assay
1,000 assays
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JumonjiC ヒストン 脱メチル化酵素およびジオキシゲナーゼ活性を簡便に検出

JumonjiC ドメインを有するヒストンリジン脱メチル化酵素  ( JMJC ) は、ヒストンリジンメチル基転移酵素活性を弱めることによりゲノムのエピジェネティックな段階を決定する極めて重要な役割を有しています。これらの酵素は、ヒストン内の特定のリジンよりメチル基を除去する「イレイサー」酵素として機能し、転写抑制または標的遺伝子の活性化に誘導します。

Succinate-Glo™ JmjC Demethylase/Hydroxylase Assay は JumonjiC ヒストン脱メチル化酵素およびFe(II)/α-ケトグルタル酸依存性デオキシゲナーゼ反応より生成するコハク酸を迅速に検出します。本アッセイでは反応産物のコハク酸を ATP に変換することでルシフェラーゼ反応による発光を生じ、この発光シグナルは基になる脱メチル化酵素 / 水酸化酵素活性に比例します。

22817166-L1-SuccinateGlo-Foreground

JmjC 脱メチル化酵素検出と Succinate-Glo™ Assay の概要

13815MA-W

The assay is performed in two steps. After the demethylase/dioxygenase reaction, Succinate Detection Reagent I is added to convert succinate to ATP and allow the newly synthesized ATP to be measured using a luciferase/luciferin reaction. The light generated correlates to the amount of succinate produced by the demethylase/dioxygenase, which indicates demethylase/dioxygenase activity.

優れた感度と直線性

13816MA-W-a

The Succinate-Glo™ Assay has good linearity and sensitivity. The assay is performed in a single well of a multiwell plate, and can be used to detect demethylase/dioxygenase activity in as little as a 5μl reaction. Light output is proportional to the concentration of succinate from low nM to 15µM.

JmjC 脱メチル化酵素活性の高感度検出

13817MA-W-a

KMD4A (BPS Bioscience Cat.# 50123) was titrated in the presence of 10μM methylated histone H3 peptide substrate (Anaspec Cat.#64452), 100μM ascorbate, 10μM Fe(II) and 10μM α-ketoglutarate.

13817MA-W-b

KDM4C (BPS Bioscience Cat.# 50105) was titrated in the presence of 10μM methylated histone H3 peptide substrate (Anaspec Cat.#64452), 100μM ascorbate, 10μM Fe(II) and 10μM α-ketoglutarate.

化合物ライブラリーを用いた脱メチル化酵素活性への効果検定

13820MA-W-a
In this experiment, KDM4C inhibitor IOX-1 (R&D Systems Cat.# 4464) was titrated in a solid white, half-volume 96-well plate in a total volume of 10μl using 110ng/well KDM4C, 10μM methylated histone H3 peptide substrate (Anaspec Cat.#64452), 100μM ascorbate, 10μM Fe(II), 10μM α-ketoglutarate and the indicated concentrations of inhibitor.

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Complete Protocol

製品仕様

You are viewing: V7990 Change Configuration

選択製品の構成品内容

Item Part # Size

Succinate, 10mM

 V897A 1 × 50μl

Succinate-Glo™ Buffer

 V847A 1 × 5ml

Succinate-Glo™ Solution

 V894A 1 × 50μl

ATP Detection Buffer

 V895A 1 × 10ml

Acetoacetyl-CoA, 100X

 V896A 1 × 100μl

ATP Detection Substrate

 V898A 1 × 1 vial

SDS

Search for SDS

分析証明書

Search by lot number

使用制限

For Research Use Only. Not for Use in Diagnostic Procedures.

保存条件

AA

パテントおよび免責条項

U.S. Pat. No. 7,700,310 and other patents and patents pending.

製品仕様

You are viewing: V7991 Change Configuration

選択製品の構成品内容

Item Part # Size

Succinate, 10mM

 V897A 1 × 50μl

Succinate-Glo™ Buffer

 V847B 1 × 50ml

Succinate-Glo™ Solution

 V894B 1 × 500μl

ATP Detection Buffer

 V895B 1 × 100ml

Acetoacetyl-CoA, 100X

 V896B 1 × 1ml

ATP Detection Substrate

 V898B 1 × 1 vial

SDS

Search for SDS

分析証明書

Search by lot number

使用制限

For Research Use Only. Not for Use in Diagnostic Procedures.

保存条件

AA

パテントおよび免責条項

U.S. Pat. No. 7,700,310 and other patents and patents pending.

技術資料

技術記事

サイテーション

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