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EpiQuik Histone Demethylase LSD1 Inhibitor Screening Assay Core Kit

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Schematic procedure for using the EpiQuik Histone Demethylase LSD1 Inhibitor Screening Assay Core Kit.
Inhibition of LSD1 by tranylcypromine (TCP) LSD1 concentration: 500 ng/well.
Input Type: Nuclear Extracts, Purified Enzyme
Research Area: Histone Methylation
Target Application: Activity Measurement
Vessel Format: 96-Well Plate
100% Guarantee: 6 months
Catalog No.SizePriceQty
P-3075A-4848 assays $163.00 
P-3075A-9696 assays $318.00 
Availability: Usually Ships In 1-2 Days 
Product Overview

The EpiQuik™ Histone Demethylase LSD1 Inhibitor Screening Assay Core Kit is a convenient set of tools that allows the experimenter to to measure inhibition of LSD1. The kit is ready-to-use and provides all the essential components needed for screening LSD1 inhibitors, which are directly interacting with LSD1 or block the binding of LSD1 to its substrate. The kit has the following advantages:

  • Fast procedure, which can be finished within 1 hour.
  • Innovative fluorescent assay without need for radioactivity, extraction, and chromatography.
  • Strip microplate format makes the assay flexible: manual or high throughput analysis.
  • Simple, reliable, and consistent assay conditions.

Note: This "core" kit does not come with enzymes included.

Principle & Procedure
The EpiQuik™ Histone Demethylase LSD1 Inhibitor Screening Assay Core Kit is designed for screening LSD1 demetylase inhibitors. In the assay with this kit, the unique di-methylated histone H3-K4 substrate is incubated with LSD1 in the strip wells. Active LSD1 binds to and demethylate histone H3-K4 substrate, producing hydrogen peroxide, which reacts with fluorogen 10-Acetyl-3,7-dihydroxy Phenoxazine and produces the highly fluorescent oxidation products. The intensity of fluorescence from oxidation products is proportional to LSD1 enzyme activity. Therefore, as LSD1 activity decreases by inhibition, the fluorescent signal decreases.

 
Fig. 1. Schematic procedure for using the EpiQuik Histone Demethylase LSD1 Inhibitor Screening Assay Core Kit.

Fig. 2. Inhibition of LSD1 by tranylcypromine (TCP) LSD1 concentration: 500 ng/well.

Product Components

HH1 (LSD1 Assay Buffer)
HH2 (LSD1 Substrate, 0.7 mM)*
HH3 (LSD1 Assay Standard, 100 mM)
HH4 (LSD1 Inhibitor, 1 mM)
HH5 (Fluoro-Developer)*
HH6 (Fluoro Enhancer)*
HH7 (Fluoro-Dilution)
8-Well Assay Strip (with Frame)
User Guide

* For maximum recovery of the products, centrifuge the original vial prior to opening the cap.

Frequently Asked Q's

1. Why do I have low activity of LSD1?
It is likely due to improper storage.

2. What settings for the fluorescence reader should I use?
For kit P-3075A, settings for the fluorescence reader should be 530ex/590em.

User Guide & MSDS

[User Guide]*
*Always use the actual User Guide that shipped with your product. Is the above file locked? You can also request user guides by emailing info@epigentek.com along with your contact information and institution name.

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