Render Target: STATIC
Render Timestamp: 2024-11-21T13:47:29.959Z
Commit: 5c4accf06eb7154018ba3f54329c7590f97f534a
XML generation date: 2024-09-30 01:58:57.391
Product last modified at: 2024-09-30T08:02:01.946Z
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PDP - Template Name: Monoclonal Antibody
PDP - Template ID: *******c5e4b77
R Recombinant
Recombinant: Superior lot-to-lot consistency, continuous supply, and animal-free manufacturing.

Phospho-Ezh2 (Thr345) (E7T7K) Rabbit mAb #45153

Filter:
  • WB
  • IP
  • ChIP
  • C&R

    Supporting Data

    REACTIVITY H M R Mk
    SENSITIVITY Endogenous
    MW (kDa) 98
    Source/Isotype Rabbit IgG
    Application Key:
    • WB-Western Blotting 
    • IP-Immunoprecipitation 
    • ChIP-Chromatin Immunoprecipitation 
    • C&R-CUT & RUN 
    Species Cross-Reactivity Key:
    • H-Human 
    • M-Mouse 
    • R-Rat 
    • Mk-Monkey 

    Product Information

    Product Usage Information

    For optimal ChIP and ChIP-seq results, use 10 μL of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.

    The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
    Application Dilution
    Western Blotting 1:1000
    Immunoprecipitation 1:50
    Chromatin IP 1:50
    Chromatin IP-seq 1:50
    CUT&RUN 1:50

    Storage

    Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

    Protocol

    Specificity / Sensitivity

    Phospho-Ezh2 (Thr345) (E7T7K) Rabbit mAb recognizes endogenous levels of total Ezh2 protein only when phosphorylated at Thr345. This antibody may detect a band of unknown origin around 135 kDa.

    Species Reactivity:

    Human, Mouse, Rat, Monkey

    Source / Purification

    Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr345 of human Ezh2 protein.

    Background

    The polycomb group (PcG) proteins are involved in maintaining the silenced state of several developmentally regulated genes and contribute to the maintenance of cell identity, cell cycle regulation, and oncogenesis (1,2). Enhancer of zeste homolog 2 (Ezh2), a member of this large protein family, contains four conserved regions including domain I, domain II, and a cysteine-rich amino acid stretch that precedes the carboxy-terminal SET domain (3). The SET domain has been linked with histone methyltransferase (HMTase) activity. Moreover, mammalian Ezh2 is a member of a histone deacetylase complex that functions in gene silencing, acting at the level of chromatin structure (4). Ezh2 complexes methylate histone H3 at Lys9 and 27 in vitro, which is thought to be involved in targeting transcriptional regulators to specific loci (5). Ezh2 is deregulated in various tumor types, and its role, both as a primary effector and as a mediator of tumorigenesis, has become a subject of increased interest (6).
    Ezh2 is phosphorylated on Thr345 by cyclin-dependent kinase 1 (CDK1), which increases targeting of Ezh2 to its target loci and promotes methylation of H3 at Lys27 (7,8). This activated form of Ezh2 may deposit initial H3K27me3 marks onto newly synthesized daughter strands of DNA during replication (9). In addition, phosphorylation of Thr345 affects Ezh2 stability by promoting its ubiquitination and subsequent degradation (9).
    For Research Use Only. Not For Use In Diagnostic Procedures.
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