5-Methylcytosine (5-mC) antibody (rAb)

Clone: RM231
Catalog No: 91311 Format: 50 µg Discontinued

Antibody Type:
Recombinant
Isotype:
IgG
Purification:
Protein A Chromatography
Host:
Rabbit
Reactivity:
Human, Not Species Specific

Applications

Methyl-DNA Immunoprecipitation Validated Dot Blot Validated Immunocytochemistry Validated

Application Notes


This product has been discontinued:
Please refer to our alternative products Catalog No. 91187, Catalog No. 39649, Catalog No. 61479
Validated Applications:
DB: 0.5 - 2 µg/ml
ICC: 0.5 - 2 µg/ml
MeDIP: 0.2 - 2 µg/ml

Immunogen

BSA-conjugated 5-methylcytosine.

Buffer

PBS with 50% glycerol, 1% BSA, and 0.09% sodium azide.

 

Dot blot of 5-Methylcytosine (5-mC) antibody (rAb).
Dot blot of double stranded DNA using 5-Methylcytosine (5-mC) antibody (RM231). The membrane was pre-spotted with 50, 5, and 0.5 ng/dot of double stranded 5-Hydroxymethylcytosine (5-hmC) DNA, 5-Methylcytosine (5-mC) DNA, and unmethylated DNA. The pre-spotted membrane was then blotted with 5-Methylcytosine (5-mC) antibody at 0.5 µg/ml.

MeDIP of 5-Methylcytosine (5-mC) antibody (rAb).
MeDIP was performed using 5-Methylcytosine (5-mC) antibody (RM231) at a 2:1 DNA:Ab ratio. 1 ng of unmethylated, 5-Methylcytosine (5-mC) or 5-Hydroxymethylcytosine (5-hmC) DNA standard (897 bp) was spiked in 1 µg of genomic DNA isolated from HeLa cells as the control. Realtime PCR was then performed to determine the capture of DNA standard as in % of input. 

Immunofluorescence stain of 5-Methylcytosine (5-mC) antibody (rAb).
HeLa cells were stained using 5-Methylcytosine (5-mC) antibody (RM231) antibody at 1 µg/ml (red). Actin filaments have been labeled with fluorescein phalloidin (green), and nuclei stained with DAPI (blue).

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.

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