Succinate-Glo™ JmjC Demethylase/Hydroxylase Assay
Sensitive, Bioluminescent Detection of Demethylase Activity
- Easy to use add-and-read assay format
- Universal assay can be used with any succinate-producing enzyme
- Optimized for screening applications; low false hits
Catalog Number:
Size
Catalog Number: V7990
Catalog Number: V7991
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。
JmjC 脱メチル化酵素検出と Succinate-Glo™ Assay の概要
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
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
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.
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
Protocols
Specifications
Catalog Number:
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 |
U.S. Pat. No. 7,700,310 and other patents and patents pending.
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 |
U.S. Pat. No. 7,700,310 and other patents and patents pending.
