Reaction Biology Corporation

Reaction Biology (RBC)社が独自に開発したHistone Methyltransferases (HMT) 、DNA Methyltransferases (DNMT) およびNucleosomes等は、RBC社が提供するプロファイリングやHTSサービスでも実際に使用されているバリデーション済みの酵素・基質です。

Nucleosomes
Methyltransferase 基質のnucleosomesにリコンビナントのnucleosomesが加わり、現在下記8種類がラインナップされています。
これらは、RIベースの³H AdoMet を用いたアッセイ(gel electrophoresis/autoradiographyやfilterplate/scintillation counting等)や、修飾部位に特異的な抗体(anti-H3K4me2等)を用いたアッセイで、ヒストンのメチル化を測定する際の基質として有用です。

Nucleosomes (HeLa Mono/Di)	Nucleosomes (HeLa Oligo)	Nucleosomes (HeLa Oligo, Biotinylated)
Recombinant Mononucleosomes H3.3	Recombinant Tetranucleosomes H3.3

《特長》

●	Each RBC HMT/DNMT is fully evaluated with a gold-standard radioisotope filtration binding assay using an array of potential substrates.
●	In addition to a simple study of enzyme activity progression curve, Michaelis-Menten kinetic parameters are also determined with at least one optimized substrate.
●	Each protein is tested with a group of commercially available inhibitors to evaluate their potencies as references.

Methyltransferase Activity of the EZH1 Complex. Methylation determined as TCA-precipitable counts in in a scintillation/filter plate assay. Reactions were 60 min., 30°C, 1 μM [3H]-SAM with 5 μM histone H3 or 0.05 mg/mL core histones.		Time courses of G9a methyltransferase reactions in the HotSpotSM assay format. G9a, at the indicated concentrations, was assayed with 0.05 mg/mL chicken core histones and 1 μM [3H]-SAM. Signal/Background ranged from 4.3 (12.5 nM G9a, 15 min.) to 51.2 (50 nM G9a, 60 min.).

Km studies Km studies can be done both for SAM and substrate

IC50 Determination of Control Compounds Each enzyme is validated by testing control compounds for each substrate.

《Available proteins and kit》 (201809更新)  NEW  Methyltransferase Enzymes Methyltransferase Substrates Reader Domain Proteins   Others Assay kit : BromoMELTSM kit Deacetylase Proteins  IDH Proteins  NQOs (QRs)  PARP  DNA Demethylation & Repair  Protein Demethylation  NUDIX & Nucleotide Pool Sanitation  Miscellaneous Apoptosis