The JARID1A Chemiluminescent Assay Kit is designed to measure
JARID1A activity for screening and profiling applications. The key to the JARID1A Chemiluminescent Assay Kit
is a highly specific antibody that recognizes demethylated substrate. With this kit, only
three simple steps on a microtiter plate are required for detection of demethylase activity.
First, JARID1A enzyme is incubated with the methylated H3 peptide for one hour. 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.
Kit comes in a convenient format, with a 96-well strip plate precoated with methylated histone H3 peptide substrate, primary antibody, the secondary HRP-labeled antibody, demethylase assay buffer, and purified JARID1A for 100 enzyme reactions.
Materials Required But Not Supplied
TBST buffer (1 x TBS, pH 8.0, containing 0.05% Tween-20) Luminometer or microplate reader capable of reading chemiluminescence Adjustable micropipettor and sterile tips Rotating or rocker platform
8-well strip plate module precoated with histone substrate
1
+4°C
UniProt #
P29375
Background
JARID1A is a member of the
Jumonji, AT-rich interactive domain 1 (JARID1) histone demethylase protein family. It
binds directly, with several other proteins, to retinoblastoma protein which regulates cell
proliferation.
References
Klose, R.J., et al. Cell 2007; 128: 889.
Storage/Stability
6 months from date of receipt, when stored as directed. Kit components require different storage conditions. Be sure to store each component at the proper temperature upon arrival.
Instructions for Use
See assay kit data sheet for detailed protocol.
Applications
Useful for studying enzyme kinetics and HTS applications.
Shipping Temperature
-80°C (dry ice)
Notes
The buffer in this kit was reformulated in May of 2015 with a different reducing agent to improve assay performance. The old formulation can still be purchased upon special request.
Contraindications
DMSO >1%, strong acids or bases, ionic detergents, high salt
Warnings
Avoid freeze/thaw cycles.
Disclaimers and Limitations
BPS Bioscience assay kits are validated using the listed components according to our specific protocol. Any deviations to kit components or protocols, such as using different proteins, cell/tissue lysates, or buffers (including using the same component from other commercial sources) will void any warranties on the performance of the assay kit and is not recommended.
End-users should immediately report any issues to [email protected] upon receipt of the kit, such as missing components, damaged vials, no dry ice, etc.