DNA demethylase (total) Activity Quantification Ultra Assay Kit (ab156908)
Key features and details
- Assay type: Quantitative
- Platform: Microplate reader
- Assay time: 4 hr
- Sample type: Cell culture extracts, Nuclear Extracts, Tissue Extracts
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