MABE176 Anti-5-hydroxymethylcytosine (5hmC) Antibody, clone AB3/63.3

MABE176
100 µg  
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      A ChIP, DB, ICC R Purified Monoclonal Antibody
      Description
      Catalogue NumberMABE176
      DescriptionAnti-5-hydroxymethylcytosine (5hmC) Antibody, clone AB3/63.3
      Alternate Names
      • 5hmeC
      • 5' hydroxymethyl cytosine
      • 5'OH methyl cytosine
      • 5'OHmeC
      • 5hmc
      • 5-hmC
      Background InformationFour types of cytosine modifications have been identified in mammalian DNA, namely 5-methylcytosine (5mC), 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC), and 5-carboxylcytosine (5caC). While 5mC, 5hmC, and 5fC occur in embryonic stem (ES) cells and many cell types in all the major organs (e.g. brain, heart, lung and kidney), 5caC has only been detected in a few cell types at a 10 times lower level than 5fC. The sequential oxidation of 5mC to 5hmC, 5fC, and 5caC catalyzed by the ten-eleven translocation (TET) enzymes has been proposed as a potential route for the demethylation of 5mC back to cytosine (C). Thymine DNA glycosylase (TDG), for example, can excise 5fC and 5caC from DNA as part of the base excision repair (BER) pathway to restore the C-nucleotide. Emerging evidences suggest that the different modifications may be selectively recognized by various proteins, and methyl-CpG-binding protein 2 (MeCP2) binding to 5hmC has been proposed as a means of epigenetic regulation in neurons. The conversion of 5hmC to 5fC, in turn, induces an enhanced binding of DNA-repair-associated proteins to 5fC, which has been proposed as a signal for DNA demethylation.
      References
      Product Information
      FormatPurified
      Control
      • Unmodified DNA, 5-Methylcytosine DNA, and 5-hydroxymethylcytosine DNA
      PresentationPurified rat monoclonal IgG2aκ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.
      Quality LevelMQ100
      Applications
      ApplicationAnti-5-hydroxymethylcytosine (5hmC) Antibody, clone AB3/63.3 is a rat monoclonal antibody that detects 5hmC modification and has been validated in Dot Blot, Immunocytochemistry, and Chromatin Immunoprecipitation (ChIP).
      Key Applications
      • Chromatin Immunoprecipitation (ChIP)
      • Dot Blot
      • Immunocytochemistry
      Application NotesChromatin Immunoprecipitation (ChIP) Analysis: A representative lot detected 5-hydroxymethylcytosine (5hmC) enrichment sites using liver chromatin preparations from wild-type female mice and mice with liver-specific Hnf4a knockout. A lower 5hmC modification level at Slc22a6 exon1 and Mgst3 intron1–2 was found associated with Hnf4a deficiency (Zhang, Q., et al. (2014). PLoS One. 9(1):e84925).

      Immunocytochemistry Analysis: A representative lot detected embryogenesis stage-dependent 5hmC immunoreactivity by indirect fluorescent staining of 4% paraformaldehyde-fixed, 0.5% Triton X-100-permeabilized, 2N HCl-DNA denatured porcine iPS and advanced pronuclear embryos of different origins (Cao, Z., et al. (2014). Theriogenology. 81(3):496-508).
      Biological Information
      Immunogen5-Hydroxymethylcytosine conjugated to BSA
      CloneAB3/63.3
      ConcentrationPlease refer to the Certificate of Analysis for the lot-specific concentration.
      HostRat
      SpecificityThis antibody detects 5-hydroxymethylcytosine, but not 5-methylcytosine or unmethylated cytosine.
      IsotypeIgG2aκ
      Species Reactivity
      • All
      Species Reactivity NoteAll. Target modification is not species-specific.
      Antibody TypeMonoclonal Antibody
      Purification MethodProtein G Purified
      Molecular WeightVariable, depending on the size(s) of the DNA molecule(s) with 5-hydroxymethylcytosine (5hmC) modification.
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceEvaluated by Dot Blot in a panel of of DNA cytosine modifications.

      Dot Blot Analysis: 1-10 µg/mL of this antibody detected 10-50 ng of DNA with 5-hydroxymethylcytosine (5hmC) modification, but not DNA with only unmodified cytosine or 5-methylcytosine (5mC) modifiction.
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsStable for 1 year at 2-8°C from date of receipt.
      Packaging Information
      Material Size100 µg
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      Anti-5-hydroxymethylcytosine (5hmC) Antibody, clone AB3/63.3 SDS

      Title

      Safety Data Sheet (SDS) 

      Anti-5-hydroxymethylcytosine (5hmC) Antibody, clone AB3/63.3 Certificates of Analysis

      TitleLot Number
      Anti-5-hydroxymethylcytosine, clone -27944812794481
      Anti-5-hydroxymethylcytosine, clone AB3/63.3 - 21611842161184
      Anti-5-hydroxymethylcytosine, clone AB3/63.3 - 22667742266774
      Anti-5-hydroxymethylcytosine, clone AB3/63.3 - 22972742297274
      Anti-5-hydroxymethylcytosine, clone AB3/63.3 - 23630512363051
      Anti-5-hydroxymethylcytosine, clone AB3/63.3 - NRG1948875NRG1948875
      Anti-5-hydroxymethylcytosine, clone AB3/63.3 -25944282594428
      Anti-5-hydroxymethylcytosine, clone AB3/63.3 Monoclonal Antibody2975844

      References

      Reference overviewPub Med ID
      Alterations of epigenetic signatures in hepatocyte nuclear factor 4α deficient mouse liver determined by improved ChIP-qPCR and (h)MeDIP-qPCR assays.
      Zhang, Q; Lei, X; Lu, H
      PloS one  9  e84925  2014

      Show Abstract
      24427299 24427299
      Dynamic reprogramming of 5-hydroxymethylcytosine during early porcine embryogenesis.
      Cao, Zubing, et al.
      Theriogenology, (2013)  2013

      Show Abstract
      24315686 24315686
      5-Hydroxymethylcytosine in the mammalian zygote is linked with epigenetic reprogramming.
      Wossidlo, Mark, et al.
      Nat Commun, 2: 241 (2011)  2011

      Show Abstract
      21407207 21407207

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      Categories

      Life Science Research > Antibodies and Assays > Primary Antibodies