Succinate-Glo™ JmjC Demethylase/Hydroxylase Assay

V7990_Succinate-Glo-JmjC DemethylaseHydroxylase Assay
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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

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Catalog number selected: V7990

$ 470.00
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Succinate-Glo™ JmjC Demethylase/Hydroxylase Assay
1,000 assays
$ 470.00
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Simple Detection of JumonjiC Histone Demethylase and Dioxygenase Activity

JumonjiC domain-containing histone lysine demethylases (JMJCs) play a pivotal role in determining the epigenetic status of the genome by counteracting the activities of histone lysine methyltransferases. These enzymes act as erasers by catalyzing the removal of methyl marks from specific lysine sites in histones, leading to either transcriptional repression or activation of target genes.

The Succinate-Glo™ JmjC Demethylase/Hydroxylase Assay rapidly detects succinate formation in JumonjiC histone demethylase and Fe(II)/α-ketoglutarate-dependent dioxygenase reactions. The assay uses the reaction product, succinate, to form ATP, which drives a bioluminescent reaction to produce a signal proportional to the original demethylase/hydroxylase activity.  

22817166-L1-SuccinateGlo-Foreground

JmjC Demethylase Detection and the Succinate-Glo™ Assay

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.

Excellent Sensitivity and Linearity

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.

Sensitive Detection of JmjC Demethylase Activity

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.

Identify Effects of Library Compounds on Demethylase Activity

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.

Protocols

Complete Protocol

Specifications

You are viewing: V7990 Change Configuration

What's in the box?

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

Certificate of Analysis

Search by lot number

Use Restrictions

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

Storage Conditions

AA

Patents and Disclaimers

U.S. Pat. Nos. 7,741,067 and 8,361,739.

Patent Pending.

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

Specifications

You are viewing: V7991 Change Configuration

What's in the box?

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

Certificate of Analysis

Search by lot number

Use Restrictions

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

Storage Conditions

AA

Patents and Disclaimers

U.S. Pat. Nos. 7,741,067 and 8,361,739.

Patent Pending.

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

Resources

Articles

Citations

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