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
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.
JmjC Demethylase Detection and the Succinate-Glo™ Assay
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
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
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.
Identify Effects of Library Compounds on Demethylase Activity
Protocols
Specifications
Catalog Number:
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 SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
U.S. Pat. No. 7,700,310 and other patents and patents pending.
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 SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
U.S. Pat. No. 7,700,310 and other patents and patents pending.
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