FBXL10 (KDM2B) Homogeneous Assay Kit

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Catalog #

50610

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Description

The FBXL10 Homogeneous Assay Kit is designed to measure the activity of the FBXL10 for screening and profiling applications. The key to the FBXL10 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 demethylase activity detection. First, a sample containing FBXL10 enzyme is incubated with the biotinylated substrate. Next, acceptor beads and primary antibody are added, then donor beads, followed by reading the Alpha-counts.

Scheme 1: Our histone demethylase assays utilize highly specific antibodies that recognize demethylated products. First, a sample containing the enzyme is incubated with a biotinylated substrate. Next, acceptor beads and primary antibody are added, then donor beads, followed by reading the Alpha-counts, as shown below.

Need us to run inhibitor screens or profile your compounds against FBXL10 (KDM2B)? Check out our Demethylase Screening Services.

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Synonyms

KDM2B, JHDM1B, CXXC2

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Product Info

Storage and Usage

Citations

Assay Kit Format

AlphaLISA®

Supplied As

Kit comes in a convenient AlphaLISA® format, with biotinylated histone H3 peptide substrate, primary antibody, methylation assay buffer, and purified FBXL10 for 96 or 384 enzyme reactions.

Materials Required But Not Supplied

AlphaLISA anti-rabbit IgG acceptor beads, 5 mg/ml (PerkinElmer #AL104L)
AlphaScreen Streptavidin-conjugated donor beads, 5 mg/ml (PerkinElmer #6760002S)
Optiplate 384 (PerkinElmer #6007290)
AlphaScreen microplate reader
Adjustable micropipettor and sterile tips

Format

96 Reactions:

Catalog Number	Component	Amount	Storage
50120	FBXL10	5 µg	-80°C	Avoid freeze/ thaw *cycles!*
52140P2	Primary antibody 16-2	5 µl	-20°C
	Biotinylated histone H3 peptide substrate	300 rxns	-80°C
79848	4x FBXL10 assay buffer 1	3 ml	-80°C
52301	4x Detection buffer	2 ml	-20°C

384 Reactions:

Catalog Number	Component	Amount	Storage
50120	FBXL10	20 µg	-80°C	Avoid freeze/ thaw *cycles!*
52140P2	Primary antibody 16-2	20 µl	-20°C
	Biotinylated histone H3 peptide substrate	500 rxns	-80°C
79848	4x FBXL10 assay buffer 1	2 ml	-80°C
52301	4x Detection buffer	2 ml	-20°C

UniProt #

Q8NHM5

Background

FBXL10, also known as KDM2B, JHDM1B, and CXXC2, is a histone lysine demethylase that exhibits demethylation activity toward trimethyl-lysine 4 (H3K4me3) and dimethyl-lysine 36 (H3K36me2) on histone H3.

References

Iwase, S., et al. Cell 2007; 128(6):1077-1088.

Storage/Stability

At least 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 protocol for detailed instructions.

Applications

Useful for studying enzyme kinetics and HTS applications.

Shipping Temperature

-80°C

Notes

AlphaScreen^® and AlphaLISA^® are registered trademarks of PerkinElmer, Inc.

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 culture medium RPMI 1640 at >1% leads to signal reduction due to the presence of excess biotin and iron in this medium. MEM, which lacks these components, does not affect AlphaScreen assays.

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.

Scientific Category

Demethylase

Regulatory Status

Research Use Only

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FBXL10 (KDM2B) Homogeneous Assay Kit

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