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DNA (Cytosine-5)-Methyltransferase 1 (DNMT1) (AA 1-125), (N-Term) antibody

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N-Term, AA 1-125
(59), (41), (35), (28), (22), (22), (19), (19), (13), (13), (13), (12), (10), (9), (9), (7), (7), (7), (6), (6), (5), (4), (3), (3), (2), (2), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1)
(389), (110), (41), (40), (36), (5), (5), (5), (3), (2), (1), (1), (1), (1), (1), (1)
(253), (111), (20), (8), (3)
(23), (23), (20), (18), (18), (18), (5), (5), (5), (5), (5), (5), (5), (5), (5), (5), (5), (5), (4), (4), (4), (4), (4), (4), (4), (4)
Western Blotting (WB), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP)
(257), (131), (104), (97), (82), (72), (46), (40), (26), (22), (11), (7), (2), (2), (1)
Pubmed 9 references available
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Quantity 0.1 mL
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Immunogen Contained within amino acids 1-125 of the N-terminus of human Dnmt1
Purification Whole antisera
Alternative Name DNMT1 (DNMT1 Antibody Abstract)
Background Inactivation of tumor suppressor genes is a key component of the development of allcommon forms of human cancers. Aberrant methylation may contribute to many of thechanges, both physiological and pathological, associated with aging, including tumordevelopment. Hypermethylation of CpG island-containing promoters is associated withthe inactivation of genes important to tumor angiogenesis and progression. DNAmethytransferase (DNMT1) is necessary for the maintenance of the methylationprocess. Alternate Names: anti-DNMT-1 antibody, anti-DNMT 1 antibody, anti-DNMT antibody, anti-DNA9cytosine-5-)-methylatransferase 1 antibody, anti-MCMT antibody, anti-DNA MTaseMmu1 antibody, anti-DNA methyltransferase M.HsaI antibody.
Gene Symbol: DNMT1
Gene ID 1786
UniProt P26358
Research Area DNA/RNA
Application Notes This Dnmt1 antibody is useful for Immunocytochemistry/Immunofluorescence, Immunoprecipitation, Immunohistochemistry on paraffin-embedded sections and Western Blot, where a band can be seen at ~190 kDa. NOTE: This antibody does not neutralize the enzymatic activity of its corresponding methyltransferase.
Recommended dilutions: Immunocytochemistry/Immunofluorescence 1:50-1:200, Immunohistochemistry 1:100, Immunohistochemistry-Paraffin 1:100, Immunoprecipitation 1:10-1:500, Western Blot 1:1000
Protocol Immunohistochemistry-Paraffin Protocol for Dnmt1 Antibody Antigen Unmasking:Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.
0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes. Staining:
1. Wash sections in deionized water three times for 5 minutes each.
. Wash sections in wash buffer for 5 minutes.
. Block each section with 100-400 µL blocking solution for 1 hour at room temperature.
. Remove blocking solution and add 100-400 µL diluted primary antibody. Incubate overnight at 4 C.
. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
. Add 100-400 µL biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
. Add 100-400 µL Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
. Wash sections three times in wash buffer for 5 minutes each.
. Add 100-400 µL DAB substrate to each section and monitor staining closely.
. As soon as the sections develop, immerse slides in deionized water.
. Counterstain sections in hematoxylin.
. Wash sections in deionized water two times for 5 minutes each.
. Dehydrate sections.
. Mount coverslips.*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.Immunocytochemistry/Immunofluorescence Protocol for Dnmt1 Antibody Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.
. Remove culture medium and add 10 % formalin to the dish. Fix at room temperature for 30 minutes.
. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
. To block nonspecific antibody binding incubate in 10 % normal goat serum from 1 hour to overnight at room temperature.
. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
Restrictions For Research Use only
Format Liquid
Preservative Without preservative
Handling Advice Avoid freeze-thaw cycles
Storage 4 °C
Storage Comment 4 °C short term. Aliquot and store at -20 °C long term.
Supplier Images
Western Blotting (WB) image for anti-DNA (Cytosine-5)-Methyltransferase 1 (DNMT1) (AA 1-125), (N-Term) antibody (ABIN151732) Western blot analysis using ABIN151732 to detect DNMT1 levels in T24 cells. Lane 1:50...
Product cited in: Morey Kinney, Smiraglia, James et al.: "Stage-specific alterations of DNA methyltransferase expression, DNA hypermethylation, and DNA hypomethylation during prostate cancer progression in the transgenic adenocarcinoma of mouse prostate ..." in: Molecular cancer research : MCR, Vol. 6, Issue 8, pp. 1365-74, 2008 (PubMed).

Estève, Chin, Smallwood et al.: "Direct interaction between DNMT1 and G9a coordinates DNA and histone methylation during replication." in: Genes & development, Vol. 20, Issue 22, pp. 3089-103, 2006 (PubMed).

Yang, Seto: "Collaborative spirit of histone deacetylases in regulating chromatin structure and gene expression." in: Current opinion in genetics & development, Vol. 13, Issue 2, pp. 143-53, 2003 (PubMed).

Bachman, Park, Rhee et al.: "Histone modifications and silencing prior to DNA methylation of a tumor suppressor gene." in: Cancer cell, Vol. 3, Issue 1, pp. 89-95, 2003 (PubMed).

Robert, Morin, Beaulieu et al.: "DNMT1 is required to maintain CpG methylation and aberrant gene silencing in human cancer cells." in: Nature genetics, Vol. 33, Issue 1, pp. 61-5, 2003 (PubMed).

Neumeister, Albanese, Balent et al.: "Senescence and epigenetic dysregulation in cancer." in: The international journal of biochemistry & cell biology, Vol. 34, Issue 11, pp. 1475-90, 2002 (PubMed).

Beaulieu, Morin, Chute et al.: "An essential role for DNA methyltransferase DNMT3B in cancer cell survival." in: The Journal of biological chemistry, Vol. 277, Issue 31, pp. 28176-81, 2002 (PubMed).

Suzuki, Gabrielson, Chen et al.: "A genomic screen for genes upregulated by demethylation and histone deacetylase inhibition in human colorectal cancer." in: Nature genetics, Vol. 31, Issue 2, pp. 141-9, 2002 (PubMed).

Rhee, Bachman, Park et al.: "DNMT1 and DNMT3b cooperate to silence genes in human cancer cells." in: Nature, Vol. 416, Issue 6880, pp. 552-6, 2002 (PubMed).

Catalog No. ABIN151732

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