JMJD2B Homogeneous Assay Kit
Catalog #50414

Size: 384 reactions

DESCRIPTION: The JMJD2B Homogeneous Assay Kit is designed to measure JMJD2B
activity for screening and profiling applications. JMJD2B is a JmjC-domain protein that
exhibits demethylation activity toward H3-K9Me3 and H3-K36Me3. The JMJD2B
Homogeneous Assay Kit comes in a AlphaLISA® format, with biotinylated histone H3
peptide substrate, primary antibody, demethylase assay buffer, and purified JMJD2B for
384 enzyme reactions. The key to the JMJD2B Homogeneous 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 methyltransferase detection. First, a sample
containing JMJD2B enzyme is incubated with the biotinylated substrate.
Next, acceptor beads and primary antibody are added, then donor beads, followed by
reading the Alpha-counts. Note: The kit is also suitable for use with cell extracts, but
nonspecific demethylase activity may be detected.

COMPONENTS:

APPLICATIONS: Great for studying enzyme kinetics and HTS applications.
 
CONTRAINDICATIONS:   Green  and blue dyes  that absorb  light  in  the AlphaScreen
signal emission  range  (520-620 nm),  such as Trypan Blue. Avoid  the use of  the potent
singlet oxygen quenchers such as sodium azide (NaN3) or metal ions (Fe2+, Fe3+, Cu2+,
Zn2+  and  Ni2+).  The  presence  of  >1%  RPMI  1640  culture medium  leads  to  a  signal
reduction  due  to  the  presence  of  excess  biotin  and  iron  in  this medium. MEM, which
lacks these components, does not affect AlphaScreen assays. 
 
STABILITY:  At least one year from date of receipt when stored as directed.

REFERENCES:  
Whetstine JR et al. Cell 2006; 125: 467. 
Fodor, B.D., Genes & Dev. 2006. 20: 1557-1562.

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