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Epigenetics and Nuclear Signaling Chromatin Binding Proteins Methylated DNA

DNA demethylase (total) Activity Quantification Ultra Assay Kit (ab156908)

DNA demethylase (total) Activity Quantification Ultra Assay Kit (ab156908)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

Key features and details

  • Assay type: Quantitative
  • Platform: Microplate reader
  • Assay time: 4 hr
  • Sample type: Cell culture extracts, Nuclear Extracts, Tissue Extracts

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Overview

  • Product name

    DNA demethylase (total) Activity Quantification Ultra Assay Kit
    See all DNA demethylase kits
  • Sample type

    Cell culture extracts, Tissue Extracts, Nuclear Extracts
  • Assay type

    Quantitative
  • Assay time

    4h 0m
  • Species reactivity

    Reacts with: Plants, Mammals
  • Product overview

    Abcam's DNA demethylase (total) Activity Quantification Ultra Assay Kit (ab156908) is a complete set of essential components which enables an experimenter to measure total DNA demethylase activity/inhibition. The kit can be used with nuclear extracts from a broad range of species such as mammals and plants, in a variety of forms including, but not limited to, cultured cells and fresh tissues.

  • Notes

    DNA demethylation is necessary for the epigenentic reprogramming of genes and is also directly involved in many important disease mechanisms such as tumor progression. Demethylation of DNA can either be passive or active, or a combination of both. Passive DNA demethylation usually takes place on newly synthesized DNA strands via DNMT1 during replication rounds. Active DNA demethylation mainly occurs by the removal of 5-methylcytosine through further modified cytosine bases which have been converted by TET enzyme-mediated oxidation. These oxidation products have been shown to be repaired by TDG, a glycosylase which is involved in base excision repair, or directly converted to cytosine by DNMT3A/ DNMT3B in oxidative states. It is proposed that DNA demethylation could also be initiated by deamination of 5-mC through candidate deaminases including AID and APOBEC1, which convert 5-mC to thymine. The resulting thymine could be repaired by BER initiated by a T-G mismatch glycosylase such as MBD4 or TDG. In addition, the 5-mC base can be directly removed in plants by the DME/ROS1 family of 5-mC DNA glycosylases, resulting in an abasic site that is repaired by the BER process.

  • Platform

    Microplate reader

Properties

  • Storage instructions

    Please refer to protocols.
  • Components 48 tests 96 tests
    10X Assay Substrate 1 x 10µl 1 x 20µl
    10X Wash Buffer 1 x 14ml 1 x 28ml
    8-Well Assay Strips (with Frame) 1 x 6 units 1 x 12 units
    Assay Control Standard, 20 µg/mL 1 x 10µl 1 x 20µl
    Binding Solution 1 x 5ml 1 x 10ml
    Capture Antibody,1000 µg/mL 1 x 4µl 1 x 8µl
    Demethylase Assay Buffer 1 x 3ml 1 x 6ml
    Detection Antibody, 400 µg/mL 1 x 6µl 1 x 12µl
    Developer Solution 1 x 5ml 1 x 10ml
    Enhancer Solution 1 x 6µl 1 x 12µl
    Stop Solution 1 x 5ml 1 x 10ml
  • Research areas

    • Epigenetics and Nuclear Signaling
    • DNA methylation
    • Methylated DNA

Images

  • Standard Curve
    Standard Curve

    Assay control standard was added into the assay wells at different concentrations and then measured with Abcam's DNA demethylase (total) Activity Quantification Ultra Assay Kit (ab156908).

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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