Recombinant CREBBP (1075-1873) protein

Catalog No: 31590 Format: 20 µg $450 Buy
Catalog No: 31990 Format: 1 mg $4,050 Buy

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Expressed In: Baculovirus Protein Species: Human

Contents

A representative Technical Data Sheet (TDS) is provided here. Please refer to the lot-specific TDS you will receive with your order for the lot-specific buffer contents and protein concentration.

Background

CREBBP (CREB Binding Protein, also known as CBP, KAT3A, RSTS) is a histone acetyltransferase (HAT) that functions primarily as a transcriptional activator. CREBBP was first isolated as a nuclear protein that specifically binds to phosphorylated cAMP-response element binding protein (CREB). It is known to play critical roles in embryonic development, growth control, and homeostasis by coupling chromatin remodeling to transcription factor recognition. It also acts as a scaffold to stabilize additional protein interactions with the transcription complex. CREBBP also acetylates non- histone proteins, like NCOA3 and FOXO1. Mutations of CREBBP cause Rubinstein-Taybi syndrome (RTS).

Application Notes

Recombinant CREBBP (1075-1873) protein is suitable for use in enzyme kinetics, inhibitor screening, and selectivity profiling.

Assay Conditions: 0.5 µg Histone H3 (Cat. No. 31294) was incubated with 0 µg (-), 0.5 µg (+) CREBBP (1075-1873) in a 20 µl reaction system containing 50 mM Tris-HCl pH 8.0, 0.1 mM EDTA, 50 ng/µl BSA, 1 mM TCEP and 20 µM Acetyl-CoA for 2 hours at room temperature. Half of each reaction was run on a 13% SDS-PAGE gel, and products were detected by Western blot. H3ac (pan-acetyl) antibody (Cat. No. 61637) was used to recognize acetylated histone H3.

Protein Details

Recombinant CREBBP (1075-1873) that includes amino acids 1075 - 1873 of human CREBBP protein (accession number NP_004371.2) was expressed in Baculovirus expression system and contains an N-terminal FLAG-Tag with a molecular weight of 94.3 kDa.

References

This product was used in the following publications:
Cell Rep (2018). Aug. 14;24(7):1722-1729. PMID: 30110629. (Histone Acetyltransferase Assay)

 

Recombinant CREBBP (1075-1873) protein, SDS-PAGE gel.Recombinant CREBBP protein was run on an 8% SDS-PAGE gel and stained with Coomassie Blue.
MW: 94.3 kDa
Purity: > 90%

Recombinant CREBBP (1075-1873) protein activity assay
50 µM H3 peptide was incubated with CREBBP (1075-1873) protein in reaction buffer for 25 min. at room temperature (RT). The reaction was stopped and developed for 15 min. in the dark at RT. Flourescence was read with excitation at 380 nm and emission at 460 nm.
HAT Assay Kit (Fluorescent, Cat.# 56100) was used for CREBBP (1075-1873) activity detection.

Recombinant CREBBP (1075-1873) protein activity assay
50 µM H3 peptide was incubated with CREBBP (1075-1873) protein in reaction buffer for 25 min. at room temperature (RT). The reaction was stopped and developed for 15 min. in the dark at RT. Flourescence was read with excitation at 380 nm and emission at 460 nm.
HAT Assay Kit (Fluorescent, Cat.# 56100) was used for CREBBP (1075-1873) activity detection.

Storage

Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.

Guarantee

This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.