Transcreener® EPIGEN SAH Methyltransferase Assay Kits
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Transcreener EPIGEN SAH 1K Assay Kit

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Technical Manual – Fluorescence Polarization (PDF)
The Transcreener® EPIGEN Methyltransferase Assay provides the following benefits:
  • Accommodates SAM concentrations ranging from 0.1 µM to 50 µM.
  • Excellent data quality (Z’ ≥0.7) and signal (≥100 mP polarization shift) at low substrate conversion using normal reaction conditions.
  • Overcomes the need for time-consuming, one-off assay development for individual members within a group transfer enzyme family by using a single set of assay reagents that detect an invariant product.
  • Far-red tracer further minimizes interference from fluorescent compounds and light scattering.

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Transcreener EPIGEN SAH Methyltransferase Assay Kit

Simple & HTS-Ready

The Transcreener® EPIGEN MT Assay is simple, HTS-ready and has the outstanding data quality and deck and signal stability that Transcreener® Assays are known for. Run your enzyme reaction, add the quenching reagent, add detection reagents (coupling enzymes are pre-combined with Transcreener® reagents), and read your plates.

How Does the Assay Work?

The Transcreener® EPIGEN Assay provides universal methyltransferase detection in an HTS-proven format. It combines the extensively validated Transcreener® AMP²/GMP² Assay with coupling enzymes that convert the SAH produced in a Methyltransferase reaction to AMP.

  • Universal: Detects any methyltransferase using any substrate
    Robust: Z’ values greater than 0.7 with low substrate consumption
  • Homogenous Format: Simple mix-and-read format for HTS workflows
  • Sensitive: Reduces enzyme usage and allows use of physiological SAM concentrations
  • >24 hr reagent and signal stability: High quality data in automated workflows at any scale
  • Validated: Optimized with a variety of enzymes including Reaction Biology’s purified MTases
Transcreener EPIGEN SAH Assay Schematic
Far Red FP Readouts Validated on Major Multimode Readers

Validation of Recombinant Human Methyltransferases with the Transcreener® EPIGEN Methyltransferase Assay

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Specifications

What’s Included

EPIGEN SAH FP Assay
1K10K
ComponentPart #QtyPart #Qty
MT Detection Mix, 10X21552 mL21562 x 10 mL
AMP2/GMP2 Antibody2114170 µL21221.7 mL
Stop Buffer A, 1X21302.5 mL21312 x 12.5 mL
5 mM SAM2117100 µL21251 mL
500 µM SAH2118250 µL21262.5 mL

What You Will Need

ComponentNotes
Methyltransferase Enzyme(s)EPIGEN assays are designed for use with purified enzyme preparations. Contaminating enzymes can produce background signal and reduce the assay window.
Enzyme Buffer ComponentsUser-supplied enzyme buffer components include enzyme, buffer, acceptor substrate, MgCl2 or MnCl2, EGTA, Brij-35, and test compounds
Assay PlatesAssay Plates Can Be Purchased Separately From BellBrook Labs.

FP - An entirely black plate with a non-binding surface is required. We suggest Corning 384-well plates (Cat. #4514) or 1536-well plates (Cat. #3728).
Plate ReaderA microplate reader configured to measure FP is required for the assay.
Liquid Handling DevicesUse liquid handling devices that can accurately dispense a minimum volume of 0.5 µL into 384-well plates.
Ultrapure WaterSome deionized water systems are contaminated with nucleases that can degrade both nucleotide substrates, and products, reducing assay performance. Careful handling and use of ultrapure water eliminates this potential problem.

Resources

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Link to: Frequently Asked Questions (FAQ’s)
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Link to: Technical Manuals
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Protocols
Link to: Publications
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Publications
Link to: Transcreener® Assays – Instrument Compatibility
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Instrument Compatibility
Link to: Application Notes
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Application Notes
Link to: Posters and Presentations
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Posters

Contact Us to Learn More

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“Some CROs reserve their best teams for high profile customers. If you’re not right there with them you get less experienced teams… BellBrook understands enzyme kinetics, so it made that part of the project so much easier. The larger CRO didn’t seem to have the expertise we needed in this area.”

“The ADP assay is a great choice for measuring activity for any ADP-generating enzyme in both real time and endpoint formats. The universal detection system has been very useful for more challenging targets.”

“Excellent blend of simplicity & good science. Excellent technical support”

“The assay itself has good reproducibility and provides a clear readout on our enzyme activity. The most important piece, however, is the level of attention to our project and rapid turnaround time. The staff at BellBrook knows what they are doing.”

“The Transcreener AMP/GMP Assay is extremely straightforward to use and is very sensitive. In a UV-based enzyme assay I needed 1 µM enzyme to see a decent signal; however using the Transcreener AMP assay I only need 10 nM. Also, BellBrook Labs have been very helpful with the assay development. Their representative has been very willing to quickly help me get my assay up and running.”

“Through the process of developing and validating an assay for HTS applications, I generated a very nice data package of robust high quality data.”

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Madison, Wisconsin 53717 USA
(608) 443-2400

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