A Homogenous, Universal Enzymatic Methyltransferase Assay for HMTs

Assay Methods

Ordering Information

Assay Services

Related Resources

AptaFluor SAH Methyltransferase Assay

AptaFluor SAH Methyltransferase Assay directly measures SAH produced by methyltransferase enzymes during methylation. These enzymes methylate a variety of substrates using S-adenosylmethionine (SAM) as the methyl donor, leaving the common product S-adenosylhomocysteine (SAH). These SAH measurements allow researchers to effectively determine the activity of the methyltransferase enzyme. The assay provides a powerful tool to screen compound libraries for methyltransferase modulators to help find new therapies for disease.

The kit comes complete with detection reagents required to measure activity. Methyltransferase enzyme is not included in the inhibitor screening assay kit. Please contact us for questions related to acquiring the enzyme.

How does the AptaFluor Methyltransferase Assay Work?

The AptaFluor SAH Methyltransferase Activity Assay determines methyltransferase enzyme activity by directly measuring SAH (S-adenosylhomocysteine) produced during the methylation of a variety of substrates. By directly detecting SAH in a homogenous format, it is possible to perform screening and enzymatic studies with virtually any methyltransferase.

AptaFluor Methyltransferase technology uses a natural occurring aptamer, or riboswitch, that selectively binds with SAH. The exquisite affinity and selectivity of the riboswitch enables screening and profiling of methyltransferases with unparalleled sensitivity.

The AptaFluor Assay is available with TR-FRET readout. It is a simple mix-and-read format. Perform your enzyme reaction, add the detection reagents, and measure. The simplicity of the system yields robust results that also makes it extremely amiable to HTS.

Direct Detection of SAH with a TR-FRET Readout

For customers who prefer TR-FRET detection. Uses the same filter set as HTRF®.

Compare Assay Sensitivity to Different Methods in this Poster

AptaFluor SAH TR-FRET Assay Tech Manual

Ultra-Sensitive

The most sensitive HTS methyltransferase assay available with an LLD of 0.6 nM SAH. This dramatically reduces enzyme usage and allows the assay to be run at or below K_m for SAM.

Easy-to-Use, Mix-and-Read, HTS-Ready

Run your enzyme reaction, add quenching reagent, add AptaFluor reagents, and read your plates. The AptaFluor Assay is compatible with 96, 384, and 1536-well formats.

AptaFluor Methyltransferase Assay Protocol Outline

Compatible with a Variety of MTase Acceptor Substrates

Toleration of MT acceptor substrates. 100 nM SAH-SAM standard curves were set up in the presence of nucleosomes (10 ng/μL), histone H3-3 (3 ng/μL), Histone H3 (1-21) peptide (10 μM) and Poly d (I-C) (2.5 mU/μL); control wells lacked a MT substrate.

Robust Assay Yields Quality Data

Excellent Z’ under a variety of assay conditions.

PRMT4 enzyme with 0.2 uM or 2 uM SAM and histone substrate as positive controls. PRMT4 and histone substrate as negative control.

Use with Physiological SAM Concentrations Lower than 100nM

SET7/9 Enzyme Titration with Linear Response Curve

Stable Detection of Methyltransferase Activity

Outstanding reagent and signal stability reduces assay variability and allows for automated workflows.

Universal – Use with Virtually Any Methyltransferase

Methyltransferase Enzyem and Substrate Chart

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Methyltransferase Assay Services

Interested in moving your program forward, but don't want to bring an assay in-house? Our scientists can help! BellBrook scientists will use their extensive biochemistry and enzymology expertise to work with you and accelerate your methyltransferase program.

Lead Discovery Services Include:

Inhibitor Screening – To identify or confirm activity with the target.
Inhibitor Potency Profiling – Dose-response with target and/or related proteins. Fast IC₅₀ results.
Residence Time Measurements – Determination of k_off using ‘jump dilution’ enzymatic assay method.
Mechanism of Action Studies – Kinetic analysis to define the mode of inhibition.

Contact Us to Learn More

Please fill out the form below. We will respond quickly to get the conservation moving and learn how we can help. We keep things discrete, confidential, and professional.

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Far Red TR-FRET Readouts Validated on Major Multimode Readers

Supplier	Instrument	TR-FRET Assays
	TriStar²S LB 942	Validated
	Mithras² LB 943	Validated

	Cytation^TM 5	Validated
	Cytation^TM 3	Validated
	Cytation^TM 1	Validated
	Synergy^TM H1	Validated
	Synergy^TM 2/H4/4	Validated
	Synergy^TM HTX	Not Capable
	Synergy^TM Neo 2	Validated

	POLARstar^® Omega	Validated
	FLUOstar^® Omega	Validated
	PHERAstar^® FSX	Validated
	PHERAstar^® Plus/FS	Validated
	CLARIOstar^® /Plus	Validated
	VANTAstar^TM	Validated

	Sense	In Review

	Analyst^® GT/HT	Validated
	Gemini^® XPS/EM	Not Capable
	SpectraMax^® M2/M2e	Not Capable
	SpectraMax^® M5/M5e/FlexStation^® 3	Validated
	SpectraMax^® Paradigm	Contact Us
	SpectraMax^® iD3/iD5	Validated

	EnVision^®/EnVision^® Xcite	Validated

	Infinite^® M1000/M1000Pro/Safire2^TM	Validated
	Infinite^® M200	Not Capable
	Infinite^® F500	Validated
	Infinite^® F200/Ultra^evolution	Contact Us
	Spark^TM 10M	Validated

What's Included


Component	Notes
SAM	SAM supplied in the kit should be used for MT enzyme reactions and to create a SAM/SAH standard curve.
SAH	SAH is used to create a SAM/SAH standard curve.
Enzyme Stop Reagent, 10X	Enzyme Stop Reagent is diluted to 1x in SAH Detection Buffer and used to quench the activity of MT enzyme components.
SAH Detection Buffer, 10X	This buffer is used to prepare the SAH Detection Mix and the Enzyme Stop Mix
P1-Biotin	P1-Biotin is a 55-base RNA oligomer with a biotin molecule at the 5' end. It is incubated with TB-Streptavidin to produce P1-Terbium, the donor piece of the SAH aptamer.
P2-Dylight 650	P2-Dylight 650 is an 18-base RNA oligomer with a Dylight 650 molecule at the 3' end. It is the acceptor piece of the SAH aptamer.
Tb-Streptavidin	Tb-Streptavidin is used to produce the Tb-labeled (donor) piece of the SAH aptamer.
Conjugation Buffer	This buffer is used for the P1-Biotin/Tb-Streptavidin conjugation reaction.

What You Will Need


Component	Notes
Methyltransferase Enzyme(s)	AptaFluor assays are designed for use with purified enzyme preparations. Contaminating enzymes can produce background signal and reduce the assay window.
Enzyme Buffer Components	User-supplied enzyme buffer components include enzyme, buffer, acceptor substrate, MgCl₂ or MnCl₂, EGTA, Brij-35, and test compounds
Assay Plates	Assay Plates Can Be Purchased Separately From BellBrook Labs. TR-FRET - An entirely white plate with a non-binding surface is required. We suggest Corning 384-well plates (Cat. #4513) or 96-well plates (Cat. #3642).
Plate Reader	A microplate reader configured to measure TR-FRET is required for the assay.
Liquid Handling Devices	Use liquid handling devices that can accurately dispense a minimum volume of 0.5 µL into 384-well plates.
Ultrapure Water	Some deionized water systems are contaminated with nucleases that can degrade both nucleotide substrates, RNA aptamers, and products, reducing assay performance. Careful handling and use of ultrapure water eliminates this potential problem.

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AptaFluor SAH Methyltransferase Assay Kits

* For custom or bulk orders (over 10,000 wells) please contact us (info@bellbrooklabs.com) for a quote.

Cat. No.	Product Description	Price	Add to Cart
3023-B	AptaFluor SAH Methyltransferase Assay (100 Assay, 96 Well)	$399.00	Add to Cart
3023-1K	AptaFluor SAH Methyltransferase Assay (1,000 Assay, 384 Well)	$1,299.00	Add to Cart
3023-10K	AptaFluor SAH Methyltransferase Assay (10,000 Assay, 384 Well)	$7,399.00	Add to Cart

Assay Plates for TR-FRET

Cat. No.	Product Description	Price	Add to Cart
2234	Corning 96-Well White Assay Plates (#3642), 3-Pack (200+ Assays)	$55.00	Add to Cart
2231	Corning 384-Well White Assay Plates (#4513), 3-Pack (1,000+ Assays)	$65.00	Add to Cart
2232	Corning 384-Well White Assay Plates (#4513), 30-Pack (10,000+ Assays)	$529.00	Add to Cart

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Methyltransferases As Drug Targets

Methyltransferases are a diverse family of enzymes that catalyze the transfer of a methyl group from S-adenosylmethionine (SAM or AdoMet), generating S-adenosylhomocysteine (SAH) as a byproduct. Methyltransferases have been associated with a variety of diseases, including cancers, inflammatory diseases, metabolic diseases, and neurodegenerative diseases (Copeland et al., 2009).

Epigenetic regulation has been shown to be a contributing factor in a variety of diseases. The discovery of methyltransferase inhibitors is currently an area of intense activity. Continuing to build an understanding of methyltransferase enzymes is step one. Finding therapeutic methods to reign in overactivity requires selective technologies that can help assess the ability to interact with the target. Robust, HTS-ready assays that are accurate yet flexible provide a powerful tool to accelerate discovery.

AptaFluor® SAH Methyltransferase Assay Kit

A Homogenous, Universal Enzymatic Methyltransferase Assay for HMTs

Assay Methods

Ordering Information

Assay Services

Related Resources

AptaFluor SAH Methyltransferase Assay

How does the AptaFluor Methyltransferase Assay Work?

Direct Detection of SAH with a TR-FRET Readout

For customers who prefer TR-FRET detection. Uses the same filter set as HTRF®.

Ultra-Sensitive

Easy-to-Use, Mix-and-Read, HTS-Ready

Compatible with a Variety of MTase Acceptor Substrates

Robust Assay Yields Quality Data

Use with Physiological SAM Concentrations Lower than 100nM

Stable Detection of Methyltransferase Activity

Universal – Use with Virtually Any Methyltransferase

Back To Top

Methyltransferase Assay Services

Lead Discovery Services Include:

Contact Us to Learn More

Back To Top

Far Red TR-FRET Readouts Validated on Major Multimode Readers

What's Included

What You Will Need

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AptaFluor SAH Methyltransferase Assay Kits

Assay Plates for TR-FRET

Back To Top

Methyltransferases As Drug Targets

Related Resources

PRMT5 is Innately Interesting

The Contribution of DNA Methyltransferase Inhibitors in Treating Myelodysplastic Syndromes

How a Methyltransferase Activity Assay Could Help Identify Novel Breast Cancer Treatments

Sensitive Methyltransferase Assay Proves Successful with PRMT4

Posters and Presentations

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