EZH2 Homogeneous Assay Kit

Catalog #

52059

$3,100 *

Size: 384 reactions

Qty

Bulk/Custom

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*US Pricing only. For international pricing, please contact your local distributor.

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Description

The EZH2 Homogenous Assay Kit is designed to measure activity of the EZH2 (enhancer of zeste homolog 2) complex (EZH2/EED/SUZ12/RbAp48/ AEBP) for screening and profiling purposes. The EZH2 Homogeneous Assay Kit comes in a convenient format 384-well AlphaLISA® format, with enough purified recombinant EZH2 complex, S-adenosylmethionine, primary antibody, substrate and buffers for 400 enzyme reactions.

The EZH2 Homogeneous Activity Assay Kit uses a highly specific antibody. In a first step, the methyltransferase is incubated with the substrate for one hour. This is followed by addition of the primary antibody and acceptor beads. Finally, the donor beads are added, and Alpha-counts are measured. The Alpha-counts are proportional to the EZH2 activity.

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Synonyms

KTM6A, EZH2, PRC2, Enhancer Of Zeste Homolog 2, EZH, EZH-2

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Product Data Gallery

Product Info

Storage and Usage

Citations

Assay Kit Format

AlphaLISA®

Materials Required But Not Supplied

AlphaLISA® Anti-Rabbit IgG Acceptor Beads, 5 mg/ml (Perkin Elmer #AL104C)
AlphaScreen® Nickel Donor Beads, 5 mg/ml (Perkin Elmer #AS101D)
Optiplate - 384 (Perkin Elmer #6007290)
AlphaScreen® microplate reader

Format

Catalog #	Name	Amount	Storage
51004	EZH2/EED/SUZ12/RbAp48/AEBP2, FLAG-Tag, His-Tag*	2 x 25 µg	-80°C
52120	20 μM S-adenosylmethionine	2 x 250 µl	-80°C
52140F	Primary Antibody 6	20 µl	-20°C
79867	Histone Octamer Substrate	400 µl	-80°C
52170-A	4x HMT Assay Buffer 2A	3 ml	-20°C
	4x Detection buffer 2	2 ml	-20°C

UniProt #

Q15910

Background

EZH2 (enhancer of zeste homolog 2) is a histone-lysine N-methyltransferase enzyme, that acts by adding methyl groups to the lysine 27 (K27) of histone H3, making it a silent chromatin. It is a functional unit of the larger complex PRC2 (polycomb repressive complex 2), which also includes EED, Suz12, AEBP2 (adipocyte enhancer-binding protein) and RbAp48 (histone-binding protein RBBP4). PRC2 is crucial for epigenetic regulation and is involved in stem cell differentiation and embryonic development. Abnormalities in PRC2 result in cancer since this complex also promotes double strand DNA repair. EZH2 is seen as an attractive target in cancer therapy, as its levels are elevated in multiple cancer types (example, breast, renal cancer, melanoma, and lymphoma). The development of inhibitors for EZH2 and PRC2 is a promising area of research for the treatment of cancer.

References

Dillon S.C., et al. 2005. Genome Biology 6:227.

Storage/Stability

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

Instructions for Use

See assay kit data sheet for detailed protocol.

Applications

Enzymatic kinetic studies and screen molecules that inhibit EHZ2 activity in drug discovery high throughput applications (HTS) applications.

Shipping Temperature

-80°C

Notes

Troubleshooting Guide
Visit bpsbioscience.com/assay-kits-faq for detailed troubleshooting instructions. For all further questions, please email [email protected]

Contraindications

The EZH2 Homogenous Assay Kit is compatible with up to 1% final DMSO concentration.
Avoid green and blue dyes that absorb light in the AlphaScreen signal emission range (λ=520-620 nm), such as Trypan Blue.
Avoid the use of potent singlet oxygen quenchers such as sodium azide (NaN₃) or metal ions (Fe²⁺, Fe³⁺, Cu²⁺, Zn²⁺ and Ni²⁺).
The presence of >1% RPMI 1640 culture medium leads to a signal reduction due to the presence of excess biotin and iron in this medium. Media like MEM, which lacks these components, does not affect AlphaScreen assays.

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

Methyltransferase

Regulatory Status

Research Use Only