Learn Why Direct ADP Detection is The Best HTS Assay Method for Kinases & ATPases

Transcreener ADP Assay Kit (FI) Method

Ordering Information

Instrument Compatibility

Related Resources

Transcreener ADP² Assay Kit - FI Readout

The Transcreener^® ADP² Assay Kit with FI readout measures ADP via antibodies selective to ADP and a fluorescent tracer. Enzymatically generated ADP displaces the tracer from an antibody-quencher conjugate, resulting in increased fluorescence. The universal detection method allows screening of virtually any kinase in a high throughput format.

The Transcreener FI ADP Assay Kit produces a simple fluorescent intensity output which can be read on even the most basic fluorescence readers typically found in academic and therapeutic research labs as well as more complex multimode plate readers. It is a simple one-step homogenous detection assay, and accommodates ATP concentrations from 0.1 to 100 µM ATP. The assay provides excellent signal at low substrate conversion, with a Z' ≥ 0.7 at 2.5% ATP conversion using 1 µM ATP.

Used in over millions of wells by pharma since 2006, Transcreener is the most extensively validated ADP detection method available backed by the best technical support in the industry.

This assay is designed to be used with purified enzyme preparations. Transcreener assays are not validated with cell lysates, blood, serum, or other biological samples.

Direct Detection of ADP to Measure Kinase Enzymatic Activity

Fluorescence Intensity

Learn More About the Transcreener ADP Assay Kit

Transcreener ADP FI Assay Tech Manual

Applications

Measure enzymatic activity of kinases
Screen compound libraries for kinase modulators
Quantify inhibitor potency
Inhibitor selectivity profiling
Measure drug-target residence time

Features

Direct detection of unlabeled ADP
Robust Assay Z' > 0.7 under initial velocity conditions
Far-red fluorescent readouts minimize compound interference
A safe, non-radioactive method
Use with ATP concentrations from 0.1 to 100 µM
Universal - use with virtually any kinase

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

Run your enzyme reaction, add Transcreener reagents, and read your plates. The ADP assay kit is compatible with 96, 384, and 1536-well formats.

Example Applications

Enzyme
ADK	RIPK2
AMPK	TBK1
IKK-beta
IRAK4
JAK1
JAK3
MAPK8
PKR
RIPK1

Detection of ADP to Measure PKR Enzymatic Activity with FI Readout

Below is an example of the Transcreener ADP² Assay with FI readout detecting ADP released by the kinase, PKR.

Back To Top

Far Red FI Readouts Validated on Major Multimode Readers

Supplier	Instrument	FI Assays
	TriStar²S LB 942	Validated
	Tristar 5	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	Validated
	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	Validated

	Analyst^® GT/HT	Validated
	Gemini^® XPS/EM	Validated
	SpectraMax^® M2/M2e	Validated
	SpectraMax^® M5/M5e/FlexStation^® 3	Validated
	SpectraMax^® Paradigm	Contact Us
	SpectraMax^® iD3/iD5	Validated
	SpectraMax^® i3x	In Review

	EnVision^®/EnVision^® Xcite	Validated

	Infinite^® M1000/M1000Pro/Safire2^TM	Validated
	Infinite^® M200	Validated
	Infinite^® F500	Validated
	Infinite^® F200/Ultra^evolution	Validated
	Spark^TM 10M	Validated

Validation Criteria for Plate Readers

384-Well Format
Z'-Factor ≥ 0.7 at 10% conversion of 10 μM ATP
Z' specifications to be met using Transcreener ADP² Assay reagents
Read time to achieve Z' specifications ≤ 5 minutes

Look for these stickers on instruments that have successfully met the validation criteria.

The instrument validation program is a collaborative effort with the major suppliers of multimode readers to insure that Transcreener assays perform optimally on your specific instrument. Scientists at BellBrook and/or the instrument vendor have tested Transcreener reagents with each instrument to determine optimal filters and settings for maximal assay performance. We provide you with application notes with all the important details, so you can be confident of success using Transcreener assays on the instrument of your choice.

Back To Top

What's Included


Component	Notes
ADP² Antibody - IRDye^® QC-1	The concentration of antibody needed for an enzyme target is dependent upon the ATP concentration and buffer conditions in the enzyme reaction. Sufficient antibody is included in the kit to complete 200 assays (Part #3013-A), 1,000 assays (Part #3013-1K), or 10,000 assays (Part #3013-10K) at an ATP concentration up to 100 µM.
ADP Alexa Fluor^® 594 Tracer	The final tracer concentration in the reaction is 4 nM.
Stop & Detect Buffer B, 10X	The Stop & Detect Buffer B components will stop enzyme reactions that require Mg²⁺. To ensure that the enzyme reaction is stopped completely, confirm that the EDTA concentration is at least equimolar to the magnesium ion concentration in the reaction. The final concentration of Stop & Detect Buffer B at the time of FI measurement is 0.5X.
ATP	The ATP supplied in this kit can be used for the enzyme reaction and to create an ADP/ATP standard curve, if desired.
ADP	ADP is used to create the ADP/ATP standard curve.

What You Will Need


Component	Notes
Kinase Enzyme	Transcreener^® ADP² Assay Kits are designed for use with purified enzyme preparations. Contaminating enzymes, such as phosphatases or nucleotidases, 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, Brij-35, and test compounds.
Assay Plates	Assay Plates Can Be Purchased Separately From BellBrook Labs. It is important to use assay plates that are entirely black with a nonbinding surface. We recommend Corning^® 384-well plates (Cat. #4514) and Corning 96-well, half-area plates (Cat. #3686).
Plate Reader	A multidetection microplate reader configured to measure FI of the Alexa Fluor^® 594 tracer is required.
Liquid Handling Devices	Use liquid handling devices that can accurately dispense a minimum volume of 2.5 µL into 384-well plates.
Ultrapure Water	Some 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.

Back To Top

Transcreener ADP² Assay Kits - FI Readout

*For custom or bulk orders (over 100,000 wells), please contact us (info@bellbrooklabs.com) for a quote.

Cat. No.	Product Description	Price	Add to Cart
3013-A	Transcreener ADP² FI Assay (200 Assay, 96 Well)	$359.00	Add to Cart
3013-1K	Transcreener ADP² FI Assay (1,000 Assay, 384 Well)	$759.00	Add to Cart
3013-10K	Transcreener ADP² FI Assay (10,000 Assay, 384 Well)	$4,299.00	Add to Cart
3013-100K	Transcreener ADP² FI Assay (10 X 10,000 Assay, 384 Well)	$21,495.00	Add to Cart

Assay Plates for FI

Cat. No.	Product Description	Price	Add to Cart
2233	Corning 96-Well Black Assay Plates (#3686), 3-Pack (200+ Assays)	$55.00	Add to Cart
2229	Corning 384-Well Black Assay Plates (#4514), 3-Pack (1,000+ Assays)	$60.00	Add to Cart
2230	Corning 384-Well Black Assay Plates (#4514), 30-Pack (10,000+ Assays)	$479.00	Add to Cart