GENTAUR Belgium BVBA BE0473327336
Voortstraat 49, 1910 Kampenhout BELGIUM
Tel 0032 16 58 90 45
GENTAUR U.S.A Genprice Inc 6017 Snell Ave, Ste 357, SanJose, CA 95123
Tel (408) 780-0908, Fax (408) 780-0908, email@example.com
Index / EpigenTek / Methylamp Global DNA Methylation Quantification Kit / Product Detail : P-1014-96 Methylamp Global DNA Methylation Quantification Kit
#P-1014-96 Methylamp Global DNA Methylation Quantification Kit
Product name : Methylamp Global DNA Methylation Quantification Kit
Catalog number : P-1014-96
Quantity: 96 assays
Supplier name : EpigenTek
Data sheet: Ask more or other datasheet now !
About this Product :Methylamp Global DNA Methylation Quantification Kit antibody storage GENTAUR recommends for long therm storage to freeze at -24 C. For short time storage up to 30 days we suggest fridge storage at 1 to 10 C. Prevent multiple freeze taw cycles of Methylamp Global DNA Methylation Quantification Kit .
More Details about
#P-1014 Methylamp Global DNA Methylation Quantification Kit will be discontinued with end of March 2012. We recommend our customers to switch to: # P-1034 MethylFlash Methylated DNA Quantification Kit (Colorimetric)
The Methylamp™ Global DNA Methylation Quantification Kit is a complete set of essential components which enables the experimenter to quantify global DNA methylation through an ELISA-like reaction using Epigentek's uniquely simplified and streamlined procedure. The entire procedure can be completed within only 4 hours in either manual or high throughput analysis. The Methylamp™ Global DNA Methylation Quantification Kit is suitable for detecting global DNA methylation status using genomic DNA isolated from variety of cultured cells, fresh and frozen tissue, paraffin-embedded tissue, plasma/serum sample, and body fluid sample, etc.
WHY CHOOSE THE METHYLAMP™ GLOBAL DNA METHYLATION QUANTIFICATION KIT?
- Very rapid procedure, which can be finished within 4 hours.
- Colorimetric quantification without radioactivity, extraction, and chromatography.
- Strip microplate format makes the assay flexible: manual or high throughput analysis.
- Simple, reliable, and consistent assay conditions.
The Methylamp™ Global DNA Methylation Quantification Kit contains all reagents required for quantification of global DNA methylation. In this assay, DNA is immobilized to the strip well specifically treated to have high affinity to the DNA. The methylated fraction of DNA can be recognized by 5-methylcytosine antibody and quantified through an ELISA-like reaction. The amount of methylated DNA is proportional to the OD intensity.
GM1 (10X Wash Buffer)
GM2 (DNA Binding Solution)
GM3 (Methylated DNA Control, 100 µg/ml)*
GM4 (Block Solution)
GM5 (Capture Antibody, 1 mg/ml)*
GM6 (Detecting Antibody, 100 µg/ml)*
GM7 (Developing Solution)
GM8 (Stop Solution)
8-Well Assay Strips (with frame)
* Spin the solution down to the bottom before use.
1. OD for blank is high, what are some suggestions to correct this?
Increase washes, extend block time to 1 hour with GU4, or decrease color development time. The stop solution should be added when the color in the positive control wells have changed to brilliant blue, and the color in blank wells have not changed, or may have only changed to slight blue. The OD for blank in general is 0.15-0.3, while OD for the positive control at 20 ng is 1-1.2. Also, increasing dilution of GU7 will help reduce background noise.
2. What is the sensitivity and DNA binding capacity of this kit?
Sensitivity limitation of the methylated DNA detection is about 0.2 ng. The reproducibility is quite good based on our quality control and customer feedback. CV% is 5-10% between wells, and 10-15% between strips, and 15-25% between different tests. The binding capacity of the well is about 400 ng of DNA.
3. Should DNA be sheared (or treated with an enzyme) prior to binding to wells?
Both un-fragmented and fragmented DNA can be tightly bound to the assay wells.
4. What is the antibody specificity of this kit?
The antibody used in the kit is monoclonal anti-5-methylcytosine Ab which recognizes any methylcytosine including 5-meC and 5-medC in CpG sites or non-cpG sites.
5. Can the kit be used for Human DNA/any model organism?
6. Can sample DNA be frozen before testing?
7. Is it necessary to know the GC content to use this assay?
No, the global DNA methylation can be quantified by comparing the amount of measured methylated DNA fraction to total DNA added for assay.
8. What are the minimum DNA quantities required for this assay?
Based on our testing, the DNA amount can be as low as 25 ng, but 50 ng DNA per each assay point is recommended.
9. Should the well be covered when incubated at 37°C for 2 hours?
No, the purpose is to dry the wells.
10. After adding stop solution, there were black precipitates in the wells.
Too strong of a reaction occurred; decrease the color development time to reduce the color reaction.
11. Kit results were fine, but OD values for samples were negative.
Increase amount of sample DNA.
12. Result for negative control gives a positive OD and there is no difference in OD between different concentrations of the positive control.
The low OD value for the positive control could be due to: 1) no binding of control DNA to the substrate coated on the strip well, and 2) capture antibody is less active. Please be sure to check the appropriate temperature and storage of the antibody and strip wells. (Capture antibody and 8 well assay strips should be stored at 4°C. Detecting antibody should be stored at -20°C.)
13. Can you explain the low OD values of my target DNA?
The meDNA fraction generally accounts for 0.8-1.5% of total DNA. Increased methylation of DNA would have higher fraction of meDNA (>2%). 200 ng of genomic DNA could contain 1.5-3 ng of meDNA and normally most of the DNA samples at this amount would give a low OD.
Barra, V. et. al. (February 2012). Bypass of cell cycle arrest induced by transient DNMT1 post-transcriptional silencing triggers aneuploidy in human cells. Cell Div. Epud ahead of print. PubMed Abstract
Liang, Y. et. al. (December 2011). Demethylation of the FCER1G promoter leads to FcεRI overexpression on monocytes of patients with atopic dermatitis. Allergy. Epub ahead of print. PubMed Abstract
Dong, R. et. al. (December 2011). Changes in epigenetic regulation of CD4+ T lymphocytesin biliary atresia. Pediatr Res. 70(6):555-9. PubMed Abstract
Sanchez-Roman. I. et. al. (December 2011). Forty percent methionine restriction lowers DNA methylation, complex I ROS generation, and oxidative damage to mtDNA and mitochondrial proteins in rat heart. J Bioenerg Biomembr. 43(6):699-708. PubMed Abstract
Kinnally, EL. et. al. (November 2011). DNA methylation as a risk factor in the effects of early life stress. Brain Behav Immun. 25(8):1548-53. PubMed Abstract
Menzel, S. et. al. (October 2011). Dissolved humic substances initiate DNA-methylation in cladocerans. Aquat Toxicol. 105(3-4):640-2. PubMed Abstract
Wang, Y. et. al. (October 2011). Intrathecal 5-azacytidine inhibits global DNA methylation and methyl- CpG-binding protein 2 expression and alleviates neuropathic pain in rats following chronic constriction injury.Brain Res. 1418:64-9. PubMed Abstract
Zeng, H. et. al. (August 2011). Dietary Selenomethionine Increases Exon-Specific DNA Methylation of the p53 Gene in Rat Liver and Colon Mucosa. J Nutr. 141(8):1464-8. PubMed Abstract
Freilinger, M. et. al. (July-August 2011). Effects of creatine supplementation in rett syndrome: a randomized, placebo-controlled trial. J Dev Behav Pediatr. 32(6):454-60. PubMed Abstract
Rudolf, E., Cervinka M. (July 2011). Stress responses of human dermal fibroblasts exposed to zinc pyrithione. Toxicol Lett. 204(2-3): 164-73. PubMed Abstract
Hervouet, E. et. al. (April 2011). Proximity ligation in situ assay for monitoring the global DNA methylation in cells. BMC Biotechnol. 11:31. PubMed Abstract
Nandakumar, V. et. al. (April 2011). Aberrant DNA hypermethylation patterns lead to transcriptional silencing of tumor suppressor genes in UVB-exposed skin and UVB-induced skin tumors of mice. Carcinogenesis. 32(4): 597-604. PubMed Abstract
Chavan-Gautam, P. et. al. (March 2011). Gestation-dependent changes in human placental global DNA methylation levels. Mol Reprod Dev. 78(3): 150. PubMed Abstract
Kulkarni, A. et. al. (March 2011). Effects of altered maternal folic acid, vitamin B12 and docosahexaenoic acid on placental global DNA methylation patterns in Wistar rats. PLoS One. 6(3): e17706. PubMed Abstract
Racila, D. et. al. (March 2011). Transient expression of OCT4 is sufficient to allow human keratinocytes to change their differentiation pathway. Gene Therapy. 18(3): 294-303. PubMed Abstract
Debien, E. et. al. (February 2011). ABT-737 and/or folate reverse the PDGF-induced alterations in the mitochondrial apoptotic pathway in low-grade glioma patients. Clinical Epigenetics. Full Text Article
Kulkarni, A. et. al. (February 2011). Global DNA Methylation Patterns in Placenta and Its Association with Maternal Hypertension in Pre-Eclampsia. DNA and Cell Biology. 30(2): 79-84. PubMed Abstract
Ward, A. et. al. (February 2011). Aberrant methylation of Polo-like kinase CpG islands in Plk4 heterozygous mice. BMC Cancer. 11:71. PubMed Abstract
Li, Y. Y. et. al. (November 2010). Lead exposure in pheochromocytoma cells induces persistent changes in amyloid precursor protein gene methylation patterns. Environmental Toxicology. Epub ahead of print. PubMed Abstract
Zhao, M. et. al. (October 2010). Abnormal DNA methylation in peripheral blood mononuclear cells from patients with vitiligo. Br J Dermatol. 163(4): 736-42. PubMed Abstract
Chen, X. et. al. (July 2010). Global DNA hypomethylation is associated with NTD-affected pregnancy: A case-control study. Birth Defects Res A Clin Mol Teratol. 88(7): 575-8. PubMed Abstract
Wan, Y. J. et. al. (July 2010). Alterations in tumor biomarker GSTP gene methylation patterns induced by prenatal exposure to PFOS. Toxicology. 274(1-3): 57-64. PubMed Abstract
Xu, N. et. al. (July 2010). Epigenetics in polycystic ovary syndrome: A pilot study of global DNA methylation. Fertil Steril. 94(2): 781-3.e1. PubMed Abstract
Zhao, M. et. al. (June 2010). Abnormal DNA methylation in peripheral blood mononuclear cells from patients with vitiligo. British Journal of Dermatology. 163(4): 736-42. PubMed Abstract
Wang, L. et .al. (May 2010). Relation between hypomethylation of long interspersed nucleotide elements and risk of neural tube defects. Am J Clin Nutr. 91(5): 1359-67. PubMed Abstract
Kastl, L. et. al. (April 2010). Effects of decitabine on expression of selected endogenous control genes in human breast cancer cells. Mol. Cell Probes. 24(2): 87-92. PubMed Abstract
Chen, J. et. al. (February 2010). Assessment of the neurotoxic mechanisms of decabrominated diphenyl ether (PBDE-209) in primary cultured neonatal rat hippocampal neurons includes alterations in second messenger signaling and oxidative stress. Toxicol Lett. 192(3): 431-9. PubMed Abstract
Batra, V. et. al. (February 2010). Enhanced one-carbon flux towards DNA methylation: Effect of dietary methyl supplements against gamma-radiation-induced epigenetic modifications. Chem Biol Interact. 183(3): 425-33. PubMed Abstract
Agrawal, A. et. al. (January 2010). Age-associated epigenetic modifications in human DNA increase its immunogenicity. Aging: Impact Journals in Biology & Medicine. 2(1): 1-8. Full PDF Article
Herbstman, J. et. al. (November 2009). Relation of DNA methylation of 5'-CpG island of ACSL3 to transplacental exposure to airborne polycyclic aromatic hydrocarbons and childhood asthma. PLoS One. 4(2): e4488. PubMed Abstract
Wang, J. et. al. (May 2009). Emerging technologies for amino acid nutrition research in the post-genome era. Amino Acids. 37(1): 177-86. PubMed Abstract
Hervouet, E. et. al. (May 2009). Folate supplementation limits the aggressiveness of glioma via the remethylation of DNA repeats element and genes governing apoptosis and proliferation. Clinical Cancer Research. 15(10): 3519-29. PubMed Abstract
Thaler, R. et. al. (March 2009). Epigenetic regulation of human buccal mucosa mitochondrial superoxide dismutase gene expression by diet. Br J Nutr. 101(5): 743-9. PubMed Abstract
Liao, Y. J. et. al. (February 2009). Characterization of a glycine N-methyltransferase gene knockout mouse model for hepatocellular carcinoma: Implications of the gender disparity in liver cancer susceptibility. International Journal of Cancer. 124(4): 816-26. PubMed Abstract
Aniagu, S. O. et. al. (February 2009). Changes in gene expression and assessment of DNA methylation in primary human hepatocytes and HepG2 cells exposed to the environmental contaminants-Hexabromocyclododecane and 17-beta oestradiol. Toxicology. 256(3): 143-51. PubMed Abstract
Hervouet, E. et. al. (November 2008). Tumor induction by disruption of the Dnmt1, PCNA and UHRF1 interactions. Nature Precedings. Full PDF Article
Luo, Y. et. al. (October 2008). Abnormal DNA methylation in T cells from patients with subacute cutaneous lupus erythematosus. Br J Dermatol. 159(4): 827-33. PubMed Abstract
Suzuki, M. et. al. (March 2008). Zebularine-induced reduction in VEGF secretion by HIF-1Î± degradation in oral squamous cell carcinoma. Molecular Medicine Reports. 1: 465-471. Full PDF Article
Novikova, S. I. et. al. (April 2008). Maternal Cocaine Administration in Mice Alters DNA Methylation and Gene Expression in Hippocampal Neurons of Neonatal and Prepubertal Offspring. PLoS ONE. 3(4): e1919. Full PDF Article
Liu, C. et. al. (February 2008). Plasma S-adenosylhomocysteine is a better biomarker of atherosclerosis than homocysteine in apolipoprotein E-deficient mice fed high dietary methionine. The Journal of Nutrition. 138(2): 311-5. PubMed Abstract
Suzuki, M. et. al. (December 2007). Epigenetic regulation of chemosensitivity to 5-fluorouracil and cisplatin by zebularine in oral squamous cell carcinoma. International Journal of Oncology. 31(6): 1449-56. Full PDF Article
Contact us about this product :
Our team will respond you as soon as possible !
52062 Aachen Deutschland
Support Karolina Elandt
Fax: (+49) 241 56 00 47 88
Logistic :0241 40 08 90 86
IBAN lautet DE8839050000107569353
Handelsregister Aachen HR B 16058
Umsatzsteuer-Identifikationsnummer *** DE 815175831
firstname.lastname@example.org | Gentaur
Genprice Inc, Logistics
547, Yurok Circle
San Jose, CA 95123
Tel (408) 780-0908,
Fax (408) 780-0908,
Genprice Inc, Invoices and accounting
6017 Snell Ave, Ste 357
San Jose, CA 95123
Canada Montreal +15149077481
Ceská republika Praha +420246019719
Finland Helsset +358942419041
Magyarország Budapest +3619980547
US New York+17185132983
WP2077: Methylation Pathways
WP704: Methylation Pathways
Related Genes :
 Aberrant DNA methylation patterns of spermatozoa in men with unexplained infertility.
 Overexpression of DNA damage-induced 45 α gene contributes to esophageal squamous cell cancer by promoter hypomethylation.
 Overexpression of the growth arrest and DNA damage-induced 45alpha gene contributes to autoimmunity by promoting DNA demethylation in lupus T cells.
Enter catalog number :