CST, 5032T, DNMT1 (D63A6) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying DNMT1. Validated for WB,IHC,IF,F,ChIP. Available in 2 sizes. Highly specific and rigorously validated in-house, DNMT1 (D63A6) Rabbit Monoclonal Antibody (CST #5032) is ready to ship. Product Usage Information For optimal ChIP results, use 10 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. Western Blotting: 1:1000 Immunohistochemistry (Paraffin): 1:50 - 1:200 Immunofluorescence (Immunocytochemistry): 1:100 Flow Cytometry (Fixed/Permeabilized): 1:50 - 1:200 Chromatin IP: 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. For a carrier free (BSA and azide free) version of this product see product # 28754 . Protocol Available protocols: Western Blotting, Immunohistochemistry (Paraffin), Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized), Chromatin IP Specificity / Sensitivity DNMT1 (D63A6) Rabbit Monoclonal Antibody detects endogenous levels of total DNMT1 protein. Species reactivity for IHC-P is human only. Species Reactivity: Human, Mouse, Rat, Monkey Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu985 of human DNMT1 protein. Background Methylation of DNA at cytosine residues in mammalian cells is a heritable, epigenetic modification that is critical for proper regulation of gene expression, genomic imprinting and development (1,2). Three families of mammalian DNA methyltransferases have been identified: DNMT1, DNMT2, and DNMT3 (1,2). DNMT1 is constitutively expressed in proliferating cells and functions as a maintenance methyltransferase, transferring proper methylation patterns to newly synthesized DNA during replication. DNMT3A and DNMT3B are strongly expressed in embryonic stem cells with reduced expression in adult somatic tissues. DNMT3A and DNMT3B function as methyltransferases that methylate previously unmethylated regions of DNA. DNMT2 is expressed at low levels in adult somatic tissues and its inactivation affects neither nor maintenance DNA methylation. DNMT1, DNMT3A, and DNMT3B together form a protein complex that interacts with histone deacetylases (HDAC1, HDAC2, Sin3A), transcriptional repressor proteins (RB, TAZ-1), and heterochromatin proteins (HP1, SUV39H1) to maintain proper levels of DNA methylation and facilitate gene silencing (3-8). Improper DNA methylation contributes to diseased states such as cancer (1,2). Hypermethylation of promoter CpG islands within tumor suppressor genes correlates with gene silencing and the development of cancer. In addition, hypomethylation of bulk genomic DNA correlates with and may contribute to the onset of cancer. DNMT1, DNMT3A, and DNMT3B are overexpressed in many cancers, including acute and chronic myelogenous leukemias, in addition to colon, breast, and stomach carcinomas (9-12). Alternate Names ADCADN; AIM; CXXC finger protein 9; CXXC-type zinc finger protein 9; CXXC9; DNA (cytosine-5-)-methyltransferase 1; DNA (cytosine-5)-methyltransferase 1; DNA methyltransferase 1; DNA methyltransferase HsaI; DNA MTase HsaI; DNMT; DNMT1; FLJ16293; HSN1E; M.HsaI; MCMT; MGC104992 Specification REACTIVITY: H M R Mk SENSITIVITY: Endogenous MW (kDa): 200 Source/Isotype: Rabbit IgG