CST, 5246L, Ezh2 (D2C9) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying EZH2. Validated for WB,WB,IP,IHC,IHC,IF,IF,F,ChIP,ChIP,C&R,C&T,eCLIP. Available in 3 sizes. Highly specific and rigorously validated in-house, Ezh2 (D2C9) Rabbit Monoclonal Antibody (CST #5246) is ready to ship. Product Usage Information For optimal ChIP and ChIP-seq results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 10 6 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 . The CUT&Tag dilution was determined using CUT&Tag Assay Kit # 77552 . Western Blotting: 1:1000 Simple Western™: 1:10 - 1:50 Immunoprecipitation: 1:300 IHC Leica Bond: 1:50 - 1:200 Immunohistochemistry (Paraffin): 1:50 Immunofluorescence (Frozen): 1:100 - 1:400 Immunofluorescence (Immunocytochemistry): 1:100 - 1:400 Flow Cytometry (Fixed/Permeabilized): 1:100 - 1:400 Chromatin IP: 1:100 Chromatin IP-seq: 1:100 CUT&RUN: 1:100 CUT&Tag: 1:100 eCLIP: 1:200 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. For a carrier free (BSA and azide free) version of this product see product # 16098 . Protocol Available protocols: Western Blotting, Immunoprecipitation, IHC Leica Bond, Immunohistochemistry (Paraffin), Immunofluorescence (Frozen), Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized), Chromatin IP, Chromatin IP-seq, CUT&RUN, CUT&Tag Specificity / Sensitivity Ezh2 (D2C9) Rabbit Monoclonal Antibody detects endogenous levels of total Ezh2 protein. This antibody does not cross-react with Ezh1 protein. Species Reactivity: Human, Mouse, Rat, Monkey Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg354 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 , 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). Alternate Names enhancer of zeste 2; enhancer of zeste 2 polycomb repressive complex 2 subunit; Enhancer of zeste homolog 2; enhancer of zeste homolog 2 (Drosophila); ENX-1; ENX1; EZH1; EZH2; EZH2b; Histone-lysine N-methyltransferase EZH2; KMT6; KMT6A; Lysine N-methyltransferase 6; MGC9169; WVS; WVS2 Specification REACTIVITY: H M R Mk SENSITIVITY: Endogenous MW (kDa): 98 Source/Isotype: Rabbit IgG