5-Methylcytosine (5-mC) antibody (pAb)

RRID: AB_2783884
Catalog No: 61255 Format: 100 µg $455 Buy
Catalog No: 61256 Format: 10 µg $110 Buy
Catalog No: 61955 Format: 50 µg $265 Buy

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Antibody Type:
Polyclonal
Isotype:
IgG
Purification:
Protein A Chromatography
Host:
Rabbit
Molecular Weight:
Not Applicable
Reactivity:
Human, Not Species Specific

Applications

ChIP Validated Methyl-DNA Immunoprecipitation Validated Dot Blot Validated Immunohistochemistry Validated

Application Notes


Validated Applications:
MeDIP: 2 - 10 µg per IP
DB: 2 µg/ml dilution

Published Applications:
ChIP
MeDIP
IHC (FFPE)

For 5-methylcytosine, we also offer AbFlex® 5-methylcytosine Recombinant Antibody (rAb).  For details, see Catalog No. 91187.

Immunogen

This 5-Methylcytosine (5-mC) antibody was raised against 5-Methyl-cytidine conjugated to KLH and recognizes 5-Methylcytosine.

Buffer

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

References

 

5-Methylcytosine antibody (pAb) tested by Methyl-DNA Immunoprecipitation (MeDIP).
Human DNA (500 ng) digested with MseI and denatured was subjected to MeDIP and then analyzed by quantitative real-time PCR with primers specific for genes that are methylated (KIAA0853, GEMIN4) or unmethylated (Untr12). The amount of MeDIP'd DNA (enriched DNA) was plotted.

5-methylcytidine antibody (pAb) tested by dot blot analysis.
DNA from the Methylated DNA Standard Kit (Catalog No. 55008) were spotted on to a positively charged nylon membrane and blotted with 5-methylcytidine antibody recognizing 5-methylcytosine (2 µg/ml dilution).

Lane 1: single stranded unmethylated DNA.
Lane 2: single-stranded DNA containing 5-methylcytosine.
Lane 3: single-stranded DNA 5-hydroxymethylcytosine.
Lane 4: double-stranded unmethylated DNA.
Lane 5: double-stranded DNA containing 5-methylcytosine.
Lane 6: double-stranded DNA containing 5-hydroxymethylcytosine.

Background

5-Methylcytosine (5-Methylcytidine) is a modified base that is found in the DNA of plants and vertebrates. DNA methylation is an epigenetic event in which DNA methyltransferases (DNMTs) catalyze the reaction of a methyl group to the fifth carbon of cytosine in a CpG dinucleotide. This modification helps to control gene expression and is also involved in genomic imprinting, while aberrant DNA methylation is often associated with disease. The 5-methylcytosine antibody has been developed to discriminate between the modified base and its normal cytosine counterpart, allowing for gene promoter methylation analysis.

qPCR Primers

Active Motif's Human Positive Control Primer Set ZC3H13 (Cat. No. 71009) and Human Negative Control Primer Set 1 (Cat. No. 71001) have been validated for performing qPCR and endpoint PCR when this antibody has been used for ChIP on human samples. For ChIP experiments with this antibody on mouse samples, the Mouse Negative Control Primer Set 1 (Cat. No. 71011) has been validated for both qPCR and endpoint PCR.

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with 5-Methylcytosine (5-mC) antibody (pAb).

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;
  • 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;
  • WB = Western Blotting