5-mC DNA ELISA Kit
D5325 / D5326
- For high-throughput detection of global 5-methylcytosine (5-mC) in DNA
- The streamlined workflow can be completed in less than 3 hours
|Detection||> 0.5% 5-mC per 100 ng DNA|
|Equipment||Incubator and ELISA plate reader. A multi-channel pipettor is recommended. An automated plate washer may be used for blocking and wash steps.|
|Input||This protocol is optimized for 100 ng input DNA/well. Compatible with DNA in the range of 10-200 ng/well.|
|Sample Source||Purified genomic DNA, plasmid DNA, PCR amplification products, or DNA fragments in water, Tris-EDTA, or similar.|
Q1: Why is there no color development after addition of HRP developer?
Q2: How to quantify the percentage of 5-mC in a DNA sample?
Q3: Can control DNAs be adjusted to low % 5-mC samples?
Q4: How should the components of the 5-mC ELISA kit be stored?
Q5: What is the shelf life of the ELISA kits?
Q6: Does the ELISA Kit detect methylated cytosine in non-CpG context?
Q7: Which wavelength should be used for the ELISA?
Q8: What is the minimum input amount of DNA per well?
DNA methylation in mouse brain tissue was quantified using the 5-mC DNA ELISA kit after exposure to different diets. Authors found that if cystathionine-β-synthase (a deficiency in this enzyme can play a role hyperhomocysteinemia) was included in the diet , 5-mC levels increased. Folic acid supplemented in the diet did not change 5-mC levels.Anuradha Kalani, Pradip K. Kamat, Srikanth Givvimani, Kasey Brown, Naira Metreveli, Suresh C. Tyagi, Neetu Tyagi. 2013. Nutri-epigenetics Ameliorates Blood–Brain Barrier Damage and Neurodegeneration in Hyperhomocysteinemia: Role of Folic Acid.
The authors showed that exposure to house-dust mites (HDM) decreased global percentage of 5-mC but increased the percentage of 5-hmC detected in the lungs of mice compared to saline treated control group.Cheng RY, Shang Y, Limjunyawong N, Dao T, Das S, Rabold R, Sham JS, Mitzner W, Tang WY. 2014. Alternation of the Lung Methylome in Allergic Airway Hyper-Responsiveness. Environmental and Molecular Mutagenesis 55:244-255.
The 5-mC DNA ELISA Kit was used to determine the effect of treament of fetal cardiomyocytes with endothelin-1 (ET-1), a protein expressed during hypoxia on methylation levels. They found that ET-1 increased %5-mC, but 5-Aza-2’-deoxycytidine blocked ET-1 effects.Alexandra Paradis, Daliao Xiao, Jianjun Zhou and Lubo Zhang. 2014. Endothelin-1 promotes cardiomyocyte terminal differentiation in the developing heart via heightened DNA methylation. Int J Med Sci.11(4):373-80.