Skip to main content
Menu
US

HTRF Anti histone H3K27 Me2-EuK mAb conjugate, 500 Assay Points

This Eu-cryptate conjugate allows specific detection of the K27Me2 motif on histone H3, as a result of methylation or demethylation enzymatic activity.

For research use only. Not for use in diagnostic procedures.

Click to copy promo code to clipboard.
SAVE 15% NOW on online orders. Use promo code below.
YEAREND15
Offer valid until 12/15. Terms and conditions apply.
Feature Specification
Application Biochemical Enzymatic Assay

This Eu-cryptate conjugate allows specific detection of the K27Me2 motif on histone H3, as a result of methylation or demethylation enzymatic activity.

For research use only. Not for use in diagnostic procedures.

Click to copy promo code to clipboard.
SAVE 15% NOW on online orders. Use promo code below.
YEAREND15
Offer valid until 12/15. Terms and conditions apply.
Product Variants
Unit Size: 500 Assay Points
Part #:
61KC2KAE
List Price
USD 1,304.00
Unit Size: 10,000 Assay Points
Part #:
61KC2KAD
List Price
USD 7,973.00

Overview

Revvity offers a line of ready to use HTRF toolbox reagents for the construction of Histone H3 methyltransferase (HMT), demethylase (HDM) and deacetylase (HDAC) activity assays. This monoclonal antibody Eu-cryptate conjugate is specific for the histone H3K27Me2 methylation mark. It can be used for the development of biochemical assays with the highest levels of specificity and sensitivity, for the screening and study of both HMTs and HDMs, such as G9a.

Specifications

Application
Biochemical Enzymatic Assay
Brand
HTRF
Detection Modality
HTRF
Molecular Modification
Methylation
Product Group
Fluorescent Reagent
Shipping Conditions
Shipped in Dry Ice
Target Class
Epigenetics
Technology
TR-FRET
Therapeutic Area
Metabolism/Diabetes
Neuroscience
Oncology & Inflammation
Unit Size
500 Assay Points

Video gallery

How it works

Assay principle

Revvity’s HTRF-based reagents give researchers the flexibility to create highly specific, custom histone modification assays for enzymes such as HMTs, DMTs or HDACs. All these assays use a biotinylated histone H3 peptide substrate, a Eu-cryptate labeled detection antibody and XL665-conjugated Streptavidin. Assay specificity is derived from the combination of the chosen Eu-cryptate labeled anti-histone antibody together with the biotinylated peptide substrate. HTRF signal is proportional to the concentration of methyl-specific peptide (e.g. unmethylated, mono-, di- or tri-methylated peptide).

HTRF Anti histone H3K9 Me1-EuK mAb conjugate 1.svg

 

Assay protocol

Two step assays are performed in a single well, require no wash steps and can easily be miniaturized to a 384-well format. The enzymatic reaction containing the biotinylated peptide, enzyme, co-factors and compounds is followed by the detection step in which Eu-labeled anti-methylated histone H3 antibody and SA-XL665 are added. The HTRF signal is detected following a brief room temperature incubation, and is stable for multiple readouts.

HTRF Anti histone H3K9 Me1-EuK mAb conjugate 2.svg

 

Assay details

Anti-histone H3 Eu-cryptate conjugates

Methylation level

K4

K9

K27

k36

Me0

V

V

 

 

Me1

V

V

V

V

Me2

V

V

V

V

Me2

V

 

V

 

 

Antibody specificity

Each Eu-cryptate labeled anti-histone antibody recognizes a single histone H3 lysine residue and methylation level. Revvity has performed cross-reactivity studies against alternative peptides and methylation levels for each antibody to ensure specificity.

PeptideMAb H3K4 H3K9 H3K27 H3K36                
  me0 me1 me2 me3 me0 me1 me2 me1 me2 me3 me1 me2
H3(1-21) 100% 1.3% 0% 0% 100% 0.7% 0% - - 0% - -
H3(21-44) 0% - - 0% - - 0% 0% 0% 0% 4.4% 0%
H3(1-21) K4Ac 0% - - - - - - - - - - -
H3(1-21) K9Ac - - - - 0.1% - - - - - - -
H3(1-21) K4me1 0% 100% 0.6% 0.5% - - 0% - - 0% - -
H3(1-21) K4me2  0% 15% 100% 7.4% - - 0% - - 0% - -
H3(1-21) K4me3 0% 0.4% 0% 100% - - 0% - - 0% - -
H3(1-21) K9me1 - - - 0% 0.4% 100% 0.1% - - 0% - -
H3(1-21) K9me2 - - - 0% 0.3% 0% 100% - - 0% - -
H3(1-21) K9me3 - - - 0.2% 0.1% 0% 0.1% - - 0.2% - -
H3(21-44) K27me1 - - - 0% - - 0.1% 100% 1.5% 0.2% - 0%
H3(21-44) K27me2 - - - 0.2% - - 0% 1.4% 100% 3.3% - 0.1%
H3(21-44) K27me3 - - - 0% - - 0.1% 0% 0% 100% - 0%
H3(21-44) K36me1 - - - 0% - - 0.1% - - 0% 100% 0.7%
H3(21-44) K36me2 - - - 0% - - 0.1% - - 0.1% 0.60% 100%
H3(21-44) K36me3 - - - 0.1% - - 0.1% - - 0.4% 0% 0%

 

Assay validation

Validation of enzymatic assays

Revvity has developed a number of HTRF assays for HMT/HDM/HDAC enzymes using the antibody conjugate epigenetic panel. The table below recaps the assay set up for H3K27 position, and provides in each case a technical note including materials and methods as well as the results obtained.

Enzyme Type Methylation level Biotinylated substrate Eu3+-Cryptate MAb  
G9a Methyl transferase K27 me0 →me1 H3(21-44)K27me0 H3K27me1  
G9a Methyl transferase K27 me0 →me2 H3(21-44)K27me0 H3K27me2  
EZH2(Y641F) Methyl transferase K27 me0 →me3 H3(1-50)K27me0 H3K27me3  

 

Resources

Are you looking for resources, click on the resource type to explore further.

1-2 of 2 Resources
Brochure Icon
Brochure
HTRF assays and reagents catalog

Discover the versatility and precision of Homogeneous Time-Resolved Fluorescence (HTRF) technology. Our HTRF portfolio offers a...

Guide Icon
Guide
HTRF solutions, guide to major applications

This guide provides you an overview of HTRF applications in several therapeutic areas.

Scroll Icon