The MethylFlash Methylated DNA Quantification Equipment (Colorimetric) is an entire set of optimized buffers and reagents to colorimetrically quantify international DNA methylation by particularly measuring ranges of 5-methylcytosine (5-mC) in an ELISA-like microplate-based format. In comparison with LUMA or LINE-1, Alu, and LTR-based assays, MethylFlash™ know-how instantly quantifies precise international DNA methylation.
This package can be particularly optimized for paired use with the MethylFlash Hydroxymethylated DNA Quantification Equipment (Colorimetric) for concurrently quantifying each methylated DNA and hydroxymethylated DNA.
The MethylFlash™ Methylated DNA Quantification Equipment (Colorimetric) is an additional refinement of our earlier Methylamp™ international DNA methylation quantification kits by simplifying the workflow and enhancing the consistency of outcomes. It makes use of an revolutionary methodology to allow background indicators to be extraordinarily low, eliminating the plate drying and blocking steps. In comparison with chromatography-based strategies comparable to HPLC or mass spectrometry, this product supplies a cheap strategy to precisely measure ranges of 5-methylcytosine. The package has the next benefits and options:
Colorimetric assay with easy-to-follow steps for comfort and velocity. The complete process might be completed inside four hours.
Revolutionary package composition allows background indicators to be extraordinarily low, which eliminates the necessity for plate blocking and permits the assay to be easy, correct, dependable, and constant.
Excessive sensitivity, of which the detection restrict might be as little as 0.2 ng of methylated DNA and accepts as little as 50 ng of enter genomic DNA.
Optimized antibody and enhancer options enable excessive specificity to 5mC, with no cross-reactivity to unmethylated cytosine and no or negligible cross-reactivity to hydroxymethylcytosine throughout the indicated focus vary of the pattern DNA.
Common optimistic and adverse controls are included, that are appropriate for quantifying methylated DNA from any species.
Strip-well microplate format makes the assay versatile: handbook or excessive throughput evaluation.
Precept & Process
The MethylFlash Methylated DNA Quantification Equipment (Colorimetric) comprises all reagents crucial for the quantification of world DNA methylation. On this assay, DNA is sure to strip wells which can be particularly handled to have a excessive DNA affinity.
The methylated fraction of DNA is detected utilizing seize and detection antibodies after which quantified by an ELISA-like response by studying the absorbance in a microplate spectrophotometer at 450 nm. The quantity of methylated DNA is proportional to the OD depth measured, which might be calculated with the package’s included formulation for relative methylation standing of two totally different DNA samples or absolute quantification of 5-methylcytosine (5mC) utilizing an ordinary curve.
Simple, Quick, and Versatile
The complete colorimetric assay has easy-to-follow steps for comfort and velocity, which might be accomplished in four hours. The strip-well microplate format permits for a versatile assay in handbook or excessive throughput evaluation. Common optimistic and adverse controls are included with the package for quantifying methylated DNA from any species comparable to mammals, vegetation, fungi, micro organism, and viruses.
Secure and Handy
All of the wanted reagents, together with adverse controls and optimistic controls, for quantification of world DNA methylation are conveniently packaged within the package. The direct colorimetric quantification of DNA samples replaces out of date or inferior strategies and eliminates the necessity for DNA digestion/denaturation, radioactivity, extraction, or chromatography.
Extremely Delicate and Particular
The novel process and proprietary package compositions enable for correct quantification of methylated DNA to be achieved with excessive sensitivity and specificity. The detection restrict of the enter DNA might be as little as 0.2 ng of methylated DNA.
Comparability of Out there ELISA-based World DNA Methylation Assay Strategies
Glorious detection restrict: 0.2 ng of 5-mC DNA
Good detection restrict: >0.5 ng of 5-mC DNA
Very poor detection restrict: >1000 ng of 5-mC DNA
>20 with very low background
<four with excessive background
No want for plate blocking and denaturation of DNA
Requires plate blocking and DNA denaturation
Requires plate blocking, DNA digestion, and antibody aggressive incubation in an extra plate
DNA Sort & Species
Common for any species, each ssDNA and dsDNA
Human and mouse ssDNA solely
Particular to 5-mC solely
Cross-reaction to 5-hmC
Each relative and absolute quantification
Secure, quantitative in absolute quantity of 5-mC, and might be universally used for any species
Unstable, can’t be quantitative in quantity, and may solely be used for human or mouse
Minimal Enter Quantity
Accuracy of Detection
Excessive, steady and quantified commonplace management. Confirmed shut correlation with LC-MS evaluation by customers
Unclear, unstable and un-quantified commonplace management
% 5-mC calculation
Very excessive revealed quotation rely
Low revealed quotation rely
Low revealed quotation rely
DNA Methylation Quantification
DNA methylation performs an essential position in regular organismal improvement and in mobile differentiation in greater organisms. Gene expression, in addition to the event of practically all kinds of most cancers, are additionally tied to DNA methylation.
For instance, international lower in 5-methylcytosine content material (DNA hypomethylation) is probably going attributable to methyl-deficiency as a consequence of quite a lot of environmental influences and has been proposed as a molecular marker in a number of organic processes comparable to most cancers. World quantification of DNA methylation is essential for understanding the roles that gene expression and silencing play within the improvement of most cancers and different ailments.