PHF8 (KDM7B) Homogeneous Assay Kit

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Catalog #

50515

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Description

The PHF8 Homogeneous Assay Kit is designed to measure PHF8activity for screening and profiling applications. PHF8, also known as JHDM1F and KDM7B, is aJumonjiC (JmjC) domain containing histone lysine demethylase that exhibits demethylationactivity toward H3-K₉Me². The PHF8 Homogeneous Assay Kit comes in a convenientAlphaLISA^® format (Scheme 1), with biotinylated histone H3 peptide substrate, primaryantibody, demethylase assay buffer, and purified PHF8 for 96 enzyme reactions. The key to thePHF8 Homogeneous Assay Kit is a highly specific antibody that recognizes demethylatedsubstrate. With this kit, only three simple steps on a microtiter plate are required formethyltransferase detection. First, a sample containing PHF8 enzyme is incubated with thebiotinylated substrate. Next, acceptor beads and primary antibody are added, then donor beads,followed by reading the Alpha-counts.

50414 scheme 1

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 above.

Need us to run inhibitor screens or profile your compounds against PHF8 (KDM7B)? Check out our Demethylase Screening Services.

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Synonyms

KDM7B, JHDM1F, PHD finger protein 8, ZNF422, AlphaLISA, Alphascreen, Alpha Lisa, Alpha screen, 50515-1, 50515-2

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Product Data Gallery

Product Info

Storage and Usage

Citations

Assay Kit Format

AlphaLISA®

Supplied As

The PHF8 Homogeneous Assay Kit comes in a convenient AlphaLISA® format (Scheme 1), with biotinylated histone H3 peptide substrate, primary antibody, demethylase assay buffer, and purified PHF8 for 96 or 384 enzyme reactions.

Materials Required But Not Supplied

AlphaLISA^® anti-RIgG acceptor beads, 5 mg/ml (PerkinElmer #AL104C)
AlphaScreen^® Streptavidin-conjugated donor beads, 5 mg/ml (PerkinElmer #6760002S)
Optiplate -96 (PerkinElmer #6005290)
AlphaScreen^® microplate reader
Adjustable micropipettor and sterile tips

Format

96 Reactions:

Catalog #	Component	Amount	Storage
50131	PHF8 (KDM7B)	20 μg	-80°C	*Avoid freeze/ thaw cycles!*
52140Q4	Primary antibody 17-4	5 μl	-80°C
	Biotinylated histone H3 peptide substrate	300 rxns	-80°C
52408	4x HDM Assay Buffer 3	3 x 1 ml	-80°C
52031	4x Detection buffer	2 ml	-20°C

384 Reactions:

Catalog #	Component	Amount	Storage
50131	PHF8 (KDM7B)	60 μg	-80°C	*Avoid freeze/ thaw cycles!*
52140Q4	Primary antibody 17-4	20 μl	-80°C
79845	Biotinylated histone H3 peptide substrate	500 rxns	-80°C
52408	4x HDM Assay Buffer 3	3 ml	-80°C
52031	4x Detection buffer	2 ml	-20°C

UniProt #

Q9UPP1

References

Horton, J.R., et al. Nature Structural & Molecular Biology 2010; 17(1): 38-43.

Storage/Stability

At least 6 months from date of receipt when stored as directed.

Instructions for Use

See product datasheet for detailed instructions.

Shipping Temperature

-80°C

Notes

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

Contraindications

Avoid 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.

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