Global 5-hmC Quantification Kit

by Active Motif
Login to view price.
55018

Description

Active Motif's Global 5-hmC Quantification Kit is an ELISA-based assay designed to detect and quantify 5-hydroxymethylcytosine (5-hmC) levels in genomic DNA from different sample types (e.g. normal and diseased), treatment conditions, clinical outcomes or environmental backgrounds. The Global 5-hmC Quantification Kit enables higher throughput processing of samples, requires only nanogram amounts of genomic DNA per well and provides results that are consistent with mass spectrometry quantification.

Assay results for Active Motif's Global 5-hmC Quantification Kit.
Assay results for Active Motif's Global 5-hmC Quantification Kit.

(Click image to enlarge)

Global 5-hmC Quantification Advantages

  • Highly sensitive; can detect as little as 0.02% 5-hmC
  • Suitable for use with tissue-derived genomic DNA
  • Can detect 5-hmC from as little as 10-50 ng of input DNA
  • Includes DNA standards for quantification of global 5-hmC levels
  • Higher throughput quantification than mass spectrometry

Hydroxymethylation is known to play a role in transcriptional regulation and embryonic development. Research has indicated it may also serve as a prognostic indicator in certain cancers and neurodegenerative disorders. Because gene-specific analysis techniques do not provide information about the global levels of DNA hydroxymethylation within a genome and mass spectrometry methods require large amounts of DNA and provide low throughput analysis, Active Motif developed the Global 5-hmC Quantification Kit. It is a simple, ELISA-based assay to detect and quantify global changes in 5-hydroxymethylcytosine levels. The colorimetric readout is easily quantified using a microplate reader at 450nm.

Active Motif’s Global 5-hmC Quantification Kit contains an optimized protocol and all necessary reagents to perform DNA fragmentation, plate coating, capture, and colorimetric detection of 5-hmC. For added convenience, DNA standards containing known levels of 5-hmC are included in the kit. Standards can be utilized to generate a standard curve for determination of % 5-hmC values for the DNA samples.

To learn more about the Global 5-hmC Quantification Kit, click on the Method, Data, or Contents tabs below. To view a manual or other related documents, click on the Documents tab below.

 
Name Format Cat No. Price  
Global 5-hmC Quantification Kit 1 x 96 rxns 55018 $430 Buy Now

How does the Global 5-hmC Quantification Kit work?

Diagram demonstrating the method of the Global 5-hmC Quantification Kit from Active Motif
Figure 1: Schematic of the Global 5-hmC Quantification Kit.

In the Global 5-hmC Quantification Kit, the genomic DNA of interest is first enzymatically digested using the provided MseI enzyme and heat denatured before addition to coated 96-well plates. Unbound DNA fragments are then washed away. A 5-hydroxymethylcytosine antibody and a secondary antibody conjugated to horseradish peroxidase (HRP) are used for detection of the hydroxymethylated fragments. The colorimetric readout is then easily quantified by spectrophotometry on a microplate reader. Results are compared to the included DNA standards to extrapolate the percent 5-hmC in the DNA sample.

Global 5-hmC Quantification Kit

The Global 5-hmC Quantification Kit is an ELISA-based assay that is designed for detection and quantitation of 5-hydroxymethylcytosine (5-hmC) levels of tissue-derived genomic DNA. DNA standards with known amounts of 5-hmC are provided for generating a standard curve to extrapolate the percentage of 5-hydroxymethylcytosine in the DNA samples.

Comparison of Active Motif's Global 5-hmC Quantification Kit and Mass Spectrometry
Figure 1: Compa

You recently viewed

Clear recently viewed