Methylflash methylated dna quantification kit

• Zachary Long
• November 15, 2021November 15, 2021
• 0

DNA Methylation Quantification

DNA methylation performs a necessary place in common organismal enchancment and in cellular differentiation in larger organisms. Gene expression, along with the occasion of virtually all types of most cancers, are moreover tied to DNA methylation.

For example, worldwide decrease in 5-methylcytosine content material materials (DNA hypomethylation) might be going attributable to methyl-deficiency as a consequence of numerous environmental influences and has been proposed as a molecular marker in quite a few natural processes similar to most cancers. World quantification of DNA methylation is crucial for understanding the roles that gene expression and silencing play throughout the enchancment of most cancers and totally different illnesses.

The MethylFlash Methylated DNA Quantification Tools (Colorimetric) is a whole set of optimized buffers and reagents to colorimetrically quantify worldwide DNA methylation by significantly measuring ranges of 5-methylcytosine (5-mC) in an ELISA-like microplate-based format. As compared with LUMA or LINE-1, Alu, and LTR-based assays, MethylFlash™ know-how immediately quantifies exact worldwide DNA methylation.

This bundle could be significantly optimized for paired use with the MethylFlash Hydroxymethylated DNA Quantification Tools (Colorimetric) for concurrently quantifying every methylated DNA and hydroxymethylated DNA.

Product Choices
The MethylFlash™ Methylated DNA Quantification Tools (Colorimetric) is an extra refinement of our earlier Methylamp™ worldwide DNA methylation quantification kits by simplifying the workflow and enhancing the consistency of outcomes. It makes use of an revolutionary methodology to permit background indicators to be terribly low, eliminating the plate drying and blocking steps. As compared with chromatography-based methods similar to HPLC or mass spectrometry, this product provides an inexpensive technique to exactly measure ranges of 5-methylcytosine. The bundle has the following advantages and choices:

• Colorimetric assay with easy-to-follow steps for consolation and velocity. The whole course of is likely to be accomplished inside 4 hours.
• Revolutionary bundle composition permits background indicators to be terribly low, which eliminates the need for plate blocking and permits the assay to be straightforward, right, reliable, and fixed.
• Extreme sensitivity, of which the detection limit is likely to 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 choices allow extreme specificity to 5mC, with no cross-reactivity to unmethylated cytosine and no or negligible cross-reactivity to hydroxymethylcytosine all through the indicated focus range of the sample DNA.
• Widespread optimistic and hostile controls are included, which can be applicable for quantifying methylated DNA from any species.
• Strip-well microplate format makes the assay versatile: handbook or extreme throughput analysis.

Principle & Course of
The MethylFlash Methylated DNA Quantification Tools (Colorimetric) includes all reagents essential for the quantification of world DNA methylation. On this assay, DNA is certain to strip wells which could be significantly dealt with to have a extreme DNA affinity.

The methylated fraction of DNA is detected using seize and detection antibodies after which quantified by an ELISA-like response by learning the absorbance in a microplate spectrophotometer at 450 nm. The amount of methylated DNA is proportional to the OD depth measured, which is likely to be calculated with the bundle’s included formulation for relative methylation standing of two completely totally different DNA samples or absolute quantification of 5-methylcytosine (5mC) using an atypical curve.

 

Easy, Fast, and Versatile
The whole colorimetric assay has easy-to-follow steps for consolation and velocity, which is likely to be completed in 4 hours. The strip-well microplate format permits for a flexible assay in handbook or extreme throughput analysis. Widespread optimistic and hostile controls are included with the bundle for quantifying methylated DNA from any species similar to mammals, vegetation, fungi, micro organism, and viruses.

Safe and Helpful
The entire wished reagents, along with hostile controls and optimistic controls, for quantification of world DNA methylation are conveniently packaged throughout the bundle. The direct colorimetric quantification of DNA samples replaces outdated or inferior methods and eliminates the need for DNA digestion/denaturation, radioactivity, extraction, or chromatography.

Extraordinarily Delicate and Explicit
The novel course of and proprietary bundle compositions allow for proper quantification of methylated DNA to be achieved with extreme sensitivity and specificity. The detection limit of the enter DNA is likely to be as little as 0.2 ng of methylated DNA.

Comparability of On the market ELISA-based World DNA Methylation Assay Methods

	MethylFlash(Colorimetric)	Competitor Z	Competitor C
Assay Principle	Direct ELISA	Direct ELISA	Indirect ELISA
Format	96-well plate	96-well plate	96-well plate
Sensitivity	Wonderful detection limit: 0.2 ng of 5-mC DNA	Good detection limit: >0.5 ng of 5-mC DNA	Very poor detection limit: >1000 ng of 5-mC DNA
S/N Ratio	>20 with very low background	<4 with extreme background	N/A*
Procedural Consolation	No need for plate blocking and denaturation of DNA	Requires plate blocking and DNA denaturation	Requires plate blocking, DNA digestion, and antibody aggressive incubation in an additional plate
Protocol Time	<4 hr	<4 hr	>eight hr
DNA Kind & Species	Widespread for any species, every ssDNA and dsDNA	Human and mouse ssDNA solely	Quite a few species
Specificity	Explicit to 5-mC solely	Cross-reaction to 5-hmC	N/A*
Quantitation Kind	Every relative and absolute quantification	Relative	Relative
Commonplace Administration	Safe, quantitative in absolute amount of 5-mC, and is likely to be universally used for any species	Unstable, cannot be quantitative in amount, and will solely be used for human or mouse	N/A*
Minimal Enter Amount	50 ng	100 ng	1000 ng
Accuracy of Detection	Extreme, regular and quantified commonplace administration. Confirmed shut correlation with LC-MS analysis by clients	Unclear, unstable and un-quantified commonplace administration	N/A*
% 5-mC calculation	Simple	Subtle	Not Supplied
Patented Methodology	Certain	No	No
Recognition	Very extreme revealed citation rely	Low revealed citation rely	Low revealed citation rely
Assist Expertise	Since 2006	Since 2013	Since 2011

 

 

 

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