5-mCpA antibody (mAb)

RRID: AB_2793764
Clone: 2C8H8A6
Aliases: Methyl-CpA, 5-methyl-cytosine-adenosine di-nucleoside, 2C8H8A6
Catalog No: 61783 Format: 100 µg $445 Buy
Catalog No: 61784 Format: 10 µg $110 Buy

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Antibody Type:
Monoclonal
Isotype:
IgG1
Purification:
Protein A Chromatography
Host:
Mouse
Reactivity:
Not Species Specific

Applications

Dot Blot Validated

Application Notes

Applications Validated by Active Motif:
DB: 1 µg/ml

Immunogen

This antibody was raised against 5-methyl-cytosine-adenosine di-nucleoside conjugated to KLH and recognized 5-mCpA. 

Buffer

Purified IgG in PBS with 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

 

5-mCpA Antibody Specificity
Dot blot analysis was used to confirm the specificity of 5-mCpA antibody for the 5-mCpA dinucleotide. Single-stranded DNA oligonucleotides (amount of oligo in nanograms listed on the left side of the blot) were spotted on to a positively charged nylon membrane and blotted with 5-mCpA (1 µg/ml dilution). Column 1: 5-meCpA. Column 2: CpA. Column 3: 5-meCpC. Column 4: 5-meCpT. Column 5: 5-meCpG. Column 6: Ap5mC

Background

DNA methylation is important for regulation of transcription, and in processes including imprinting, gene silencing and cancer development. Methylation occurs predominantly at cytosine within the dinucleotide CpG (meCpG), which is frequently found in promoter regions near transcription start sites, as well as in promoters for functional non-coding RNAs.  However, methylation at CpG dinucleotides makes them susceptible to both spontaneous deamination and enzyme-mediated deamination, resulting in thymine substitution (T/G mismatch) and the formation of a CpA dinucleotide in the opposite strand.  Therefore, there is a strong correlation of CpG dinucleotide depletion or “suppression” with an observed increase in TpG/CpA dinucleotides.
In mammals, there are certain cell types in which significant levels of methylation at CpA, (meCpA), CpT (meCpT), CpC (meCpC) is also observed, including embryonic stem cells, oocytes, primordial germ cells and neurons.  Early observations suggest that meCpA, in particular, has different nuclear distribution than meCpG, and that meCpA may associate with active transcription rather than suppression. 

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • ChIP = ChIP Sequencing;
  • CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
  • CUT&Tag = Cleavage Under Targets and Tagmentation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • NEU = Neutralizing;
  • WB = Western Blotting