EZH2/EED/SUZ12/RbAp48, His-tag, FLAG-tag Recombinant

Catalog #

51099

$820 *

Size: 50 µg

Qty

Bulk/Custom

Add to Favorites

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

●

Purchase

●

Description

Complex of human EZH2 (GenBank Accession No. NM_004456), a.a. 2-end with N-terminal His-tag, MW= 86 kDa, human EED (NM_003797), a.a. 2-end with N-terminal FLAG-tag, MW= 51 kDa, human SUZ12 (NM_015355), a.a. 2-end with N-terminal His-tag, MW = 87 kDa, and human RbAp48 (NM_005610), a.a. 2-end with N-terminal His-tag, MW = 48 kDa, co-expressed in a baculovirus expression system.

●

Synonyms

KTM6A, EZH2, PRC2, Enhancer Of Zeste Homolog 2; RBAP48, Retinoblastoma Binding Protein 4 (RBBP4), NURF55, Chromatin Assembly Factor 1 Subunit C, CAF1 p48; Embryonic Ectoderm Development, hEED, EED, WAIT-1; SUZ12, Suppressor Of Zeste 12 Homolog, CHET9, JJA

●

Product Data Gallery

4-20% Gradient SDS-PAGE Coomassie Staining

Specific Activity

Product Info

Storage and Usage

Citations

Species

Human

Host Species/Expression System

Sf9 insect cells

Purity

≥90%

Format

Aqueous buffer solution

Formulation

45 mM Tris-HCl, pH 8.0, 124 mM NaCl, 2.4 mM KCl, 3 mM DTT, 225 mM imidazole, and 10% glycerol.

272 kDa (complex)

Amino Acids

2–end (all components)

Specific Activity

0.046 pmol/min/Âµg

Genbank #

NM_004456 (EZH2), NM_003797(EED), NM_015355 (SUZ12), NM_005610(RbAP48)

UniProt #

Q15910

Tag(s)

N-terminal His-tag (SUZ12, EZH2, RbAP48), N-terminal FLAG-tag (EED)

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. Varambally, S. et al. Science 322 (5908), 1695-1699 (2008).
2. Rakotobe, D. et al., Virol. J. 5, 32 (2008).

Storage/Stability

>6 months at -80°C.

Assay Conditions

50 µl reaction mix (50 mM Tris pH 8.8, 5 mM MgCl₂, 4 mM DTT, 20 µM S-adenosylmethionine, and 50-200 ng enzyme complex) add to the wells coated with the substrate. Incubate for 1 hour. Add antibody against methylated K27 residue of histone H3, incubate 1 hour. Then incubate 1 hour with the HRP-labeled secondary antibody, followed by addition of the HRP substrate to produce chemiluminescence.

Instructions for Use

Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.

Applications

Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

Shipping Temperature

-80°C

Warnings

Avoid freeze/thaw cycles

Scientific Category

Methyltransferase

Regulatory Status

Research Use Only

Skip to the end of the images gallery

Skip to the beginning of the images gallery

●

Datasheets/Protocols

51099_Lot110901.pdf

Datasheets provide example lot-specific information. Contact us to request information on currently available lots.

● General Protocols

● FAQs

● Contact Technical Service

●

EZH2/EED/SUZ12/RbAp48, His-tag, FLAG-tag Recombinant

PRODUCTS

SERVICES

RESOURCES

COMPANY