EZH2 (Y641N) TR-FRET Assay Kit

Catalog #

52078

$1,550 *

Size: 384 reactions

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Description

The EZH2 (Y641N) TR-FRET Assay Kit is designed to measure activity of the mutant EZH2 complex [EZH2 (Y641N)/EED/SUZ12/RbAp48/AEBP] in a homogeneous 384 reaction format. This FRET-based assay requires no time-consuming washing steps, making it especially suitable for high throughput screening applications. The key to the EZH2 (Y641N) Assay Kit is a highly specific antibody that recognizes methylated Histone H3K27. With this kit, only three simple steps on a microtiter plate are required for methyltransferase detection. First, Sadenosylmethionine is incubated with a sample containing assay buffer and methyltransferase enzyme for 4 hours. Next, antibody to methylated H3K27 is added. Finally, dye-labeled acceptor is added followed by fluorescence detection.

This product has been cited 1 time.

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Synonyms

HTRF

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Product Data Gallery

Product Info

Storage and Usage

Citations1

Assay Kit Format

TR-FRET

Supplied As

The EZH2 (Y641N) TR-FRET Assay Kit comes in a convenient format, with histone H3 peptide substrate, a Tb-labeled antibody against methylated K27 residue of Histone H3, S-adenosylmethionine, methyltransferase assay buffer, TR-FRET detection buffer, dye-lab

Format

Catalog #	Component	Amount	Storage
51028	EZH2 (Y641N)/EED/SUZ12/ RbAp48/AEBP2	100 µg	-80°C	Avoid freeze/ thaw cycles!
52120	20 µM S-adenosylmethionine	2 x 250 µl	-80°C
	Tb-labeled antibody	20 µl	-80°C
	Biotinylated histone H3 peptide substrate*	500 reactions	-80°C
52170-A	4x HMT Assay Buffer 2A	3 ml	-20°C
	Dye-labeled acceptor	2 x 10 µl	-20°C
	TR-FRET Detection Buffer	4 ml	-20°C
79969	White, Nonbinding Corning, low volume, microtiter plate	1	Room Temp

*Resuspend in 500 µl of distilled water

UniProt #

Q15910

Background

EZH2 is the catalytic subunit of the PRC2/EED-EZH2 complex, which methylates Lys9 (H3K9me) and Lys27 (H3K27me) of histone H3, leading to transcriptional repression of the affected target gene. Able to mono-, di- and trimethylate Lys27 of histone H3 to form H3K27me1, H3K27me2 and H3K27me3, respectively. Compared to EZH2-containing complexes, it is more abundant in embryonic stem cells and plays a major role in forming H3K27me3, which is required for embryonic stem cell identity and proper differentiation. The PRC2/EED-EZH2 complex may also serve as a recruiting platform for DNA methyltransferases, thereby linking two epigenetic repression systems. Genes repressed by the PRC2/EED-EZH2 complex include HOXC8, HOXA9, MYT1, CDKN2A and retinoic acid target genes. EZH2 can also methylate non-histone proteins such as the transcription factor GATA4.

References

1. Dillon, S.C., et al. Genome Biology 2005; 6:227
2. Morin, R.D., et al. Nat Genet. 2010, 42(2):181

Storage/Stability

At least 6 months from date of receipt when stored as directed.

Instructions for Use

See datasheet for detailed protocol.

Shipping Temperature

-80°C

Notes

MATERIALS OR INSTRUMENTS REQUIRED BUT NOT SUPPLIED:
Fluorescent microplate reader capable of measuring Time Resolved Fluorescence Resonance Energy Transfer (TR-FRET)
Adjustable micropipettor and sterile tips

This mutation is often cited in the scientific literature as Y641N in variant 3 (GenBank Accession No. NM_001203247).

Warnings

Avoid freeze/thaw cycles.

Disclaimers and Limitations

BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.

End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.

Scientific Category

Methyltransferases

Regulatory Status

Research Use Only

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EZH2 (Y641N) TR-FRET Assay Kit

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