P300 Homogeneous Assay Kit

Catalog #

50078

$1,135 *

Size: 384 reactions

Qty

Bulk/Custom

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*US Pricing only. For international pricing, please contact your local distributor.

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Description

The p300 Homogeneous Assay Kit contains an AlphaLISA^® assay designed for screening inhibitors of p300 in a convenient 384-well format. This efficient method requires no time-consuming washing steps. Only a few steps are needed. The assay is based on the enzymatic transfer of an acetyl group from acetyl CoA to a specific lysine residue within a biotinylated peptide substrate. After incubation with p300, the sample is incubated with a combination of an antibody that specifically binds the acetylated peptide and acceptor beads that bind the antibody. This is followed by incubation with streptavidin-labeled donor beads and reading of the Alpha-counts.

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Synonyms

EP300, KAT3B, RSTS2, E1A binding protein, cbp

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Product Data Gallery

Product Info

Storage and Usage

Citations

Assay Kit Format

AlphaLISA®

Supplied As

The p300 Homogeneous Assay Kit comes in a convenient AlphaLISA^® format, with biotinylated histone substrate, Acetyl CoA, primary antibody, assay buffers, and purified p300 for 384 enzyme reactions.

Materials Required But Not Supplied

Anti-Rabbit AlphaLISA® Acceptor Beads, 5 mg/ml (PerkinElmer #AL104)
AlphaScreen® Streptavidin-conjugated Donor Beads, 5 mg/ml (PerkinElmer #6760002)
Optiplate -384 (PerkinElmer #6007290)
AlphaScreen® microplate reader
Adjustable micropipettor and sterile tips

Format

Catalog #	Name	Amount	Storage
50071	Human recombinant P300 enzyme*	1 µg	-80°C
	Acetyl CoA (0.1 mM)	10 μl	-20°C
	Biotinylated H3 Peptide Substrate	400 μl	-80°C
79708	2X HAT assay buffer	10 ml	-20°C
	0.5 M DTT	200 μl	-20°C
79706	Primary Antibody 327	40 μl	-80°C
52301	4X Detection Buffer 1	2 ml	-80°C

*The concentration of p300 is lot-specific and will be indicated on the tube containing the enzyme.

UniProt #

Q09472

Background

Histone acetyltransferase p300, also known as p300 HAT or EP300, is a histone acetyltransferase that regulates gene transcription through chromatin remodeling. Therefore, p300 plays an important role in the regulation of cell proliferation or differentiation. p300 is closely related to CREB binding protein. It interacts with many transcription factors. It also mediates cAMP-gene regulation by binding specifically to the phosphorylated CREB protein. Mutations in p300 are involved in the rare genetic disease Rubinstein–Taybi syndrome and in several types of cancer, including stomach, colon, breast, and pancreatic cancer.

References

Trievel, R. C., et al. (2000). Anal. Biochem. 287(2):319-28.

Storage/Stability

This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.

Applications

Study enzyme kinetics of p300
Screen small molecular inhibitors for drug discovery and High throughput (HTS) applications.

Notes

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

Contraindications

DMSO concentrations above 0.5%.
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^®

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

Acetyltransferase

Regulatory Status

Research Use Only

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P300 Homogeneous Assay Kit

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