Product nameMethylated DNA Immunoprecipitation (MeDIP) Kit - DNA
See all 5-methylcytosine kits
Assay time3h 00m
Species reactivityReacts with: Other species, Mammals
DNA methylation is a covalent modification of the cytosine ring at the 5' position of a CpG dinucleotide which leads to epigenetic inactivation of genes when found in 5'-CpG-3'dinucleotides within promoters or in the first exon of genes. It has been demonstrated that DNA methylation plays an important role in the regulation of gene expression, tumorigenesis, and other genetic and epigenetic diseases, and that alterations in DNA methylation patterns are associated with cancer.
Methylated DNA Immunoprecipitation (MeDIP) Kit - DNA (ab117133) enables the user to enrich methylated DNA by using an antibody specific to methyl cytosine (5-mC) to immunoprecipitate methylated genomic DNA (MeDIP). The enriched methylated fractions can then be used in various downstream applications including qualitative and quantitative PCR, as well as southern blot and DNA microarray.
NOTE: the size of this kit is based on number of TESTS not SAMPLES. A test simply refers to a single assay well. If you are not sure which size is most suitable for you, contact our Scientific Support Team.
Storage instructionsStore at +4°C. Please refer to protocols.
Components 24 tests 48 tests 8-Well Assay Strips (with Frame) 3 units 6 units 8-Well Strip Caps 3 units 6 units Anti-5-Methylcytosine (1 mg/mL) 1 x 25µl 1 x 50µl Antibody Buffer 1 x 8ml 1 x 16ml Binding Buffer 1 x 5ml 1 x 8ml DNA Release Buffer 1 x 2ml 1 x 4ml Elution Buffer 1 x 0.6ml 1 x 1.2ml F-Collection Tube 30 units 50 units F-Spin Column 30 units 50 units Normal Mouse IgG (1 mg/mL) 1 x 10µl 1 x 20µl Proteinase K (10 mg/mL) 1 x 25µl 1 x 50µl Reaction Buffer 1 x 4ml 1 x 8ml Wash Buffer 1 x 16ml 2 x 16ml
MeDIP was performed using 1μg of DNA with ab117133. Real-time PCR was carried out with MeDIP DNAs and the frequency of DNA methylation of immunoprecipitated DNA vs input DNA was calculated.
Captured methylated DNA was used for analyzing methylation level of GAPDH and MLH1 promoter with the use of primers and probes specific to GAPDH and MLH1 promoters, respectively.
For enrichment of methylated DNA using ab117133, DNA (0.5 ug) isolated from MCF-7 cells was added into the microwell. Methylated DNA was captured by 5-mC antibody prebound to the microwells.
This product has been referenced in:
- Han B et al. Variation of DNA Methylome of Zebrafish Cells under Cold Pressure. PLoS One 11:e0160358 (2016). MeDIP . Read more (PubMed: 27494266) »
- Schultz BM et al. Enhancers compete with a long non-coding RNA for regulation of the Kcnq1 domain. Nucleic Acids Res 43:745-59 (2015). MeDIP ; Mouse . Read more (PubMed: 25539921) »