EMARS reagent|東京未来スタイルは研究用試薬の輸入商社です。

EMARS Reagent “Ar-Flu”

 

Comprehensively labeling of cell surface molecular clustering in living cells

EMARS (Enzyme-Mediated Activation of Radical Sources) is a labeling method that labels molecules comprehensively within a limited distance (300nm) from the probed molecules on which HRP is set. The EMARS method proposes to be a powerful tool to elucidate molecular clustering on the cell surface of living cells in various contexts.

 

EMARS reagent and EMARS reaction

EMARS method

First step: setting HRP on cell membrane

Second step: EMARS reaction (a case of setting HRP by antibody)

Schematic diagram of the in vivo EMARS method Living cells are treated with HRP-conjugated cognitive molecules, and subsequently treated with EMARS reagent which consist of aryl azide compound bonded with labeling compound. Aryl azide the part of EMARS reagent is converted to active radical species by HRP and attack the molecules located within a limited distance from HRP set on a given molecule on the cell surface and label the coclustered molecules.　After the EMARS reaction, the membrane proteins are solubilized and the labeled molecules by EMARS reagents are identified by using ordinary method of protein analysis such as antibody array.

 

Last step: analysis of FITC labeled molecule

 

→ Product list

 

Product specifications sheet

Application protocols
in preparation

→ Reference

[1] Jiang,S., Kotani,N., Honke,K., et.al., A proteomics approach to the cell-surface interactome using the enzyme-mediated activation of radical sources reaction. Proteomics 2012,12,54-62

[2] Taniguchi,N.,Commentary-Searching for partners, Proteomics 2012,12,9-10

 

EMARS Reagent “Ar-Flu”

 

Comprehensively labeling of cell surface molecular clustering in living cells

EMARS (Enzyme-Mediated Activation of Radical Sources) is a labeling method that labels molecules comprehensively within a limited distance (300nm) from the probed molecules on which HRP is set. The EMARS method proposes to be a powerful tool to elucidate molecular clustering on the cell surface of living cells in various contexts.

 

EMARS reagent and EMARS reaction

EMARS method

First step: setting HRP on cell membrane

Second step: EMARS reaction (a case of setting HRP by antibody)

Schematic diagram of the in vivo EMARS method Living cells are treated with HRP-conjugated cognitive molecules, and subsequently treated with EMARS reagent which consist of aryl azide compound bonded with labeling compound. Aryl azide the part of EMARS reagent is converted to active radical species by HRP and attack the molecules located within a limited distance from HRP set on a given molecule on the cell surface and label the coclustered molecules.　After the EMARS reaction, the membrane proteins are solubilized and the labeled molecules by EMARS reagents are identified by using ordinary method of protein analysis such as antibody array.

 

Last step: analysis of FITC labeled molecule

 

→ Product list

 

Product specifications sheet

Application protocols
in preparation

→ Reference

[1] Jiang,S., Kotani,N., Honke,K., et.al., A proteomics approach to the cell-surface interactome using the enzyme-mediated activation of radical sources reaction. Proteomics 2012,12,54-62

[2] Taniguchi,N.,Commentary-Searching for partners, Proteomics 2012,12,9-10

 

生細胞膜上の分子間相互作用を簡便に解析する研究用試薬

 

EMARS試薬 “Ar-Flu”とは？

EMARS試薬Ar-Fluは、EMARS法 (Enzyme-Mediated Activation of Radical Sources method)と名付けられた、生細胞の細胞膜上に存在する標的分子の近傍(200-300nm)に存在する分子を網羅的に標識する方法に利用できる研究用試薬です。
EMARS法によって、細胞膜上に存在する生体機能分子の「会合」現象とその結果生じる機能分子の集合体の役割を解明することが可能となります。

 

EMARS試薬及びEMARS反応

 

 

HRPによりEMARS反応が起きる範囲が細胞膜上のマイクロドメインの大きさに一致し、また、HRPが細胞に無害であるため、細胞膜上マイクロドメインでの分子間相互作用を生細胞上で観察することが出来る。

 

EMARS法の概要

 

Step. 1 標的分子をHRPで標識する

 

Step. 2 EMARS反応（HRP標識抗体）

①生細胞に、HRP標識された認識分子（抗体など）を添加し、さらにアリールアジドと標識試薬の化合
物であるEMARS試薬を添加する。
②EMARS試薬を構成するアリールアジドが、HRPの存在下でラジカル化され、細胞表面の限られた範
囲（HRPの近傍300nm以内）に集まっている分子を攻撃し、それらの会合分子を標識する。
③EMARS反応の後、 EMARS試薬で標識された分子を抗体アレイやその他のタンパク解析の手法で
同定する。

 

Step. 3 FITCで標識された分子をMS解析等で解析する

 

 

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→ 参考文献・資料

[1] Jiang,S., Kotani,N., Honke,K., et.al., A proteomics approach to the cell-surface interactome using the enzyme-mediated activation of radical sources reaction. Proteomics 2012,12,54-62 [2] Taniguchi,N.,Commentary-Searching for partners, Proteomics 2012,12,9-10