m6A RNA甲基化定量试剂盒价格_品牌:Epigentek-丁香通官网

供应商：上海研卉生物科技有限公司

英文名：EpiQuik M6A RNA Methylation Quantification Kit （Colorimetric）（48 reactions）

规格：48 reactions

m6A RNA甲基化定量试剂盒
EpiQuik M6A RNA Methylation Quantification Kit （Colorimetric）（48 reactions）
随着全球科学家们在RNA甲基化研究的不断深入，对相关产品的需求愈加显著。为更好的支持RNA甲基化（m6A）研究，世界顶级表观遗传学供应商Epigentek
m6A RNA定量试剂盒（P-9005)产品说明：
EpiQuik M6A RNA Methylation Quantification Kit（比色法）是一组优化的、完整的的试剂组合，可以通过比色的方法定量RNA中N6-甲基腺嘌呤（m6A）。它可以直接检测哺乳动物，植物，真菌细菌和病毒的总RNA样本的m6A的甲基化程度。

Input Type:	RNA
Research Area:	RNA Methylation
Target Application:	Amount Quantitation
Vessel Format:	96-Well Plate

The EpiQuik™ m6A RNA Methylation Quantification Kit (Colorimetric) is a complete set of optimized buffers and reagents to colorimetrically quantify N6-methyladenosine (m6A or 6mA) in RNA. It is suitable for a direct detection of m6A RNA methylation status using total RNA isolated from any species such as mammals, plants, fungi, bacteria, and viruses. The kit has the following advantages and features:

Colorimetric ELISA-like assay using an m6A antibody with easy-to-follow steps for convenience and speed. The entire procedure can be completed within 3 hours and 45 minutes.
High sensitivity, of which the detection limit can be as low as 10 pg of m6A.
Unique binding solution allows RNA >70 nts to be tightly bound to the wells, which enables quantification of m6A from both mRNA and ncRNA such as tRNA, rRNA, and snRNA.
Optimized antibody and enhancer solutions allow high specificity to m6A, with no cross-reactivity to unmethylated adenosine within the indicated concentration range of the sample RNA.
Universal positive and negative controls are included, which are suitable for quantifying m6A from any species.
Strip-well microplate format makes the assay flexible for manual or high throughput analysis.
Simple, reliable, and consistent assay conditions.

Background Information
N6-methyladenosine (m6A) is the most common and abundant modification in RNA molecules present in eukaryotes. The m6A modification is catalyzed by a methyltransferase complex METTL3 and removed by the recently discovered m6A RNA demethylases FTO and ALKBH5, which catalyze m6A demethylation in an α-ketoglutarate (α-KG)- and Fe2+-dependent manner. It was shown that METTL3, FTO, and ALKBH5 play important roles in many biological processes, ranging from development and metabolism to fertility. m6A accounts for more than 80% of all RNA base methylations and exists in various species. m6A is mainly distributed in mRNA and also occurs in non-coding RNA such as tRNA, rRNA, and snRNA. The relative abundance of m6A in mRNA transcripts has been shown to affect RNA metabolism processes such as splicing, nuclear export, translation ability and stability, and RNA transcription. Abnormal m6A methylation levels induced by defects in m6A RNA methylase and demethylase could lead to dysfunction of RNA and diseases. For example, abnormally low levels of m6A in target mRNAs due to increased FTO activity in patients with FTO mutations, through an as-yet undefined pathway, contributes to the onset of obesity and related diseases. The dynamic and reversible chemical m6A modification in RNA may also serve as a novel epigenetic marker of profound biological significance. Therefore, more useful information for a better understanding of m6A RNA methylation levels and distribution on RNA transcripts could benefit diagnostics and therapeutics of disease.

Principle & Procedure
In this assay, total RNA is bound to strip wells using a RNA high binding solution. N6-methyladenosine (m6A) is detected using a specific capture N6-methyladenosine antibody and detection antibody. The detected signal is enhanced and then quantified colorimetrically by reading the absorbance in a microplate spectrophotometer at a wavelength of 450 nm. The amount of m6A is proportional to the OD intensity measured. Both negative and positive RNA controls are provided in this kit. A standard curve can be performed (range: 0.02 to 1 ng of m6A) or a single quantity of m6A can be used as a positive control. Because m6A content can vary from tissue to tissue, and from normal and diseased states, or vary under treated and untreated conditions, it is advised to run replicate samples to ensure that the signal generated is validated. This kit will allow the user to quantify an absolute amount of m6A and determine the relative m6A RNA methylation states of two different RNA samples.

Starting Materials
The input material required is RNA, which can be isolated from various tissue or cell samples such as cells from flask or microplate cultured cells, fresh and frozen tissues, paraffin-embedded tissues, blood, body fluid samples, etc. The amount of RNA for each assay can be 100 ng to 300 ng. For optimal quantification, the input RNA amount should be 200 ng, as the abundance of m6A is generally less than 0.1% of total RNA.

WB (10X Wash Buffer)
BS (Binding Solution)
NC (Negative Control, 100 µg/ml)*
PC (Positive Control, m6A 2 µg/ml)*
CA (Capture Antibody, 1000 X) *
DA (Detection Antibody, 1000 X)*
ES (Enhancer Solution)*
DS (Developer Solution)
SS (Stop Solution)
8-Well Assay Strips (With Frame)
User Guide

Product Citations

Zheng ZQ et. al. (July 2019). Long non-coding RNA FAM225A promotes nasopharyngeal carcinoma tumorigenesis and metastasis by acting as ceRNA to sponge miR-590-3p/miR-1275 and upregulate ITGB3. Cancer Res.

Xu Y et. al. (July 2019). The N6-methyladenosine mRNA methylase METTL14 promotes renal ischemic reperfusion injury via suppressing YAP1. J Cell Biochem.

Liu T et. al. (June 2019). Dysregulated N6-methyladenosine methylation writer METTL3 contributes to the proliferation and migration of gastric cancer. J Cell Physiol.

Kwon J et. al. (June 2019). Functional roles of hnRNPA2/B1 regulated by METTL3 in mammalian embryonic development. Sci Rep. 9(1):8640.

Liu Q et. al. (June 2019). METTL3 promotes experimental osteoarthritis development by regulating inflammatory response and apoptosis in chondrocyte. Biochem Biophys Res Commun.

Cayir A. et. al. (May 2019). Exposure to environmental toxicants reducesglobal N6-methyladenosine RNA methylation and alters expression of RNA methylation modulator genes Environmental Research.

Su H et. al. (May 2019). Transcriptome-wide map of m6A circRNAs identified in hypoxic pulmonary hypertension rat model bioRxiv.

Heng J et. al. (May 2019). Maternal heat stress regulates the early fat deposition partly through modification of m6A RNA methylation in neonatal piglets. Cell Stress Chaperones. 24(3):635-645.

Zhu H et. al. (April 2019). ALKBH5 inhibited autophagy of epithelial ovarian cancer through miR-7 and BCL-2. J Exp Clin Cancer Res. 38(1):163.

Zhao T et. al. (April 2019). Oxidative stress: One potential factor for arsenite-induced increase of N6-methyladenosine in human keratinocytes. Environ Toxicol Pharmacol. 69:95-103.

Fukumoto T et. al. (April 2019). N6-methylation of adenosine (m6A) of FZD10 mRNA contributes to PARP inhibitor resistance. Cancer Res.