Recombinant KAT2A (GCN5) protein

Catalog No: 31591 Format: 20 µg $475 Buy
Catalog No: 31991 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

GCN5 (General Control Of Amino Acid Synthesis Protein 5, also known as KAT2A, Lysine Acetyltransferase 2A) is a histone acetyltransferase (HAT) that functions primarily as a transcriptional activator. It also acts as a repressor of NF-kappa-B by promoting ubiquitination of the NF-kappa-B subunit RELA in a HAT- independent manner. KAT2A (GCN5) has significant histone acetyltransferase activity with core histones, but not with nucleosome core particles. It also acetylates non-histone proteins, such as CEBPB. KAT2A (GCN5) is a core component of the ATAC complex, which is a complex with histone acetyltransferase activity on histones H3 and H4.

Application Notes

Recombinant KAT2A (GCN5) protein is suitable for use in enzyme kinetics, inhibitor screening, and selectivity profiling.

Assay Conditions: 0.5 µg Histone H3.1 (Cat. No. 31294) was incubated with 0 µg (-), 0.1 µg (+), 0.2 µg (++), 0.4 µg (+++) GCN5 protein in a 30 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. All samples were run on a 12.5% 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 KAT2A (GCN5) protein was expressed in a Baculovirus expression system as the full length protein (accession number NP_066564.2) with an N-terminal FLAG-Tag. The molecular weight of the protein is 95.6 kDa.

 

Recombinant KAT2A (GCN5) protein, SDS-PAGE gel
Recombinant KAT2A (GCN5) protein was run on an 8% SDS-PAGE gel and stained with Coomassie Blue.
MW: 95.6 kDa
Purity: > 85%

Recombinant KAT2A (GCN5) protein activity assay
50 µM H3 peptide was incubated with KAT2A (GCN5) protein in reaction buffer for 25 minutes at room temperature. The reaction was stopped and developed for 15 minutes in the dark at RT. Fluorescence was read with excitation at 380 nm and emission at 460 nm.
HAT Assay Kit (Fluorescent, Cat# 56100) was used for KAT2A (GCN5) activity detection.

Recombinant KAT2A (GCN5) protein activity assay
50 µM H3 peptide was incubated with KAT2A (GCN5) protein in reaction buffer for 25 minutes at room temperature. The reaction was stopped and developed for 15 minutes in the dark at RT. Fluorescence was read with excitation at 380 nm and emission at 460 nm.
HAT Assay Kit (Fluorescent, Cat# 56100) was used for KAT2A (GCN5) 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.