ALKBH5 Chemiluminescent Assay Kit
The ALKBH5 Chemiluminescent Assay Kit is designed to measure ALKBH5 activity for screening and profiling applications. The key to the ALKBH5 Chemiluminescent Assay Kit is a highly specific antibody that recognizes methylated substrate. Signal is inversely related to ALKBH5 demethylase activity. With this kit, only three simple steps on a microtiter plate are required for detection of demethylase activity. First, ALKBH5 enzyme is incubated with the substrate. Next, primary antibody is added. Finally, the plate is treated with an HRP-labeled secondary antibody followed by the addition of the HRP substrate to produce chemiluminescence that can be measured using a chemiluminescence reader.
Need us to run inhibitor screens or profile your compounds against ALKBH5? Check out our Demethylase Screening Services or DNA Replication and Repair Screening Services.
TBST buffer (1x TBS, pH 8.0, containing 0.05% Tween-20), prepared with DEPC-treated water
Luminometer or microplate reader capable of reading chemiluminescence
Adjustable micropipettor and sterile tips
Rotating or rocker platform
Catalog # | Component | Amount | Storage | |
100057 | ALKBH5 | 100 µg | -80°C | Avoid freeze/ thaw cycles! |
52140Z4 | Primary antibody 29 | 100 µl | -80°C | |
52131H | Secondary HRP-labeled antibody 2 | 10 µl | -80°C | |
4x ALKBH5 assay buffer | 3 x 1 m | -80°C | ||
79556 | Blocking buffer 1 | 50 ml | +4°C | |
79670 | ELISA ECL substrate A (transparent bottle) |
6 ml | Room Temp | |
ELISA ECL substrate B (brown bottle) |
6 ml | Room Temp | ||
White 8-well strip plate module precoated with RNA substrate* | 1 | +4°C |
*NOTE: It is critical to perform all steps of the assay using RNAse-free conditions.
ALKBH5 (AlkB Homolog 5, RNA Demethylase) is an enzyme that removes N6-Methyladenosine (m6A) from target mRNAs. ALKBH5 has been connected to various cancers, viral replication, and metabolic diseases while also being shown to remove the alkylation damage and restore adenine in DNA.
Zheng, G., Dahl, J.A., et al. 2013. ALKBH5 Is a Mammalian RNA Demethylase that Impacts RNA Metabolism and Mouse Fertility, Molecular Cell, 49(1):18-29.