GSH-Glo™ Glutathione Assay

GSH-Glo™ Glutathione Assayは、細胞内あるいは様々な生体サンプルからグルタチオン（GSH）を定量するための発光セルベースアッセイシステムです。グルタチオンレベルの変化は毒性反応の評価や酸化ストレス（アポトーシスや細胞死へ移行する可能性を有す）の指標として重要です。本アッセイは、GSH存在下でルシフェリン誘導体がルシフェリンに変換される反応がベースとなっています。反応はキットに添付されるグルタチオンS-トランスフェラーゼ（GST）酵素により触媒されます。生成したルシフェリンは、グロータイプの発光を生じるUltra-Glo™ Recombinant Luciferaseを用いた反応により検出され、生じたシグナルは細胞内に存在するグルタチオン量に比例します。このアッセイ法は、比色法や蛍光法に変わるシンプル、迅速で高感度な方法で、ハイスループットなアプリケーションにも容易に適応します。

迅速：30分で結果が得られます
シンプルな測定法：2種類の試薬を加えるだけなので、従来のGSHアッセイ法に比べ、ステップ数が少なく、容易にハイスループットアプリケーションに適応させることができます（除タンパク質ステップも不要）
高感度：発光法は他法で起こる蛍光による特有のバックグラウンドの問題を回避できるため、優れたシグナル/ノイズ比を実現
安定なシグナル：発光半減期は5時間以上

細胞死の指標として細胞内あるいは組織抽出液中のGSHレベルを測定
細胞、組織あるいは血液ライセートのGSHレベルを変動させる薬剤や新規化学物質のスクリーニング

【パフォーマンス】製品概説

Want to measure the ratio of reduced to oxidized glutathione?

The GSH/GSSG-Glo™ Assay is a luminescent assay that can quantify total glutathione (GSH + GSSG), GSSG and GSH-to-GSSG ratios in cultured cells. Learn more.

The GSH-Glo™ Glutathione Assay

6956ma-w: Overview of Luminescent GSH-Glo™ Glutathione Assay

The GSH assay is performed in two steps. In the first step, cells are lysed in the presence of the luciferin-NT substrate and glutathione S-transferase. Glutathione in the cells drives the formation of luciferin. In the 2nd step, Luciferin Detection Reagent is added to produce light that is directly proportional to the amount of GSH in the reaction.

GSH Depletion and Recovery in Treated Cells

7330ma-w: GSH depletion in HeLa cells treated with a GSH synthesis inhibitor. — HeLa cells (5000 cells/well in 96 well format) were exposed to l-Buthionine-sulfoximine (BSO). BSO inhibits GSH synthesis, thus reducing cellular GSH level.

**BSO + Recovery:** Cells treated with BSO for 20 hours, washed 2X with PBS and covered with fresh media without BSO. Cells assayed for GSH after 40 hours.

**20hr BSO:** Cells treated with BSO and assayed for GSH after 20 hours.

**40hr BSO:** Cells treated with BSO, washed at 20 hours and fresh media plus BSO added. Cells assayed for GSH after 40 hours.

The simple two-reagent-addition GSH detection assay minimizes the number of assay steps compared to conventional GSH assays and is easily adapted to higher throughput applications. No deproteination step is required, and results are achieved in as quickly as 30 minutes.

The luminescent method of the GSH-Glo™ Glutathione Assay avoids inherent background fluorescence associated with other methods, thereby providing excellent signal-to-background ratios. A half-life greater than 5 hours results in a stable signal, enabling batch processing of many plates at once using the GSH-Glo™ Assay.

The GSH-Glo™ Assay is recommended for measurement of total glutathione levels in samples. To measure the ratio, we recommend the GSH/GSSG-Glo™ Assay.

GSH Depletion by Acetaminophen in Hepatocytes

7364ma-w: GSH levels assessed in liver cells treated with acetaminophen.

GSH levels in lysates from adherent cells in 24 well plates were measured with the GSH-Glo™ Glutathione Assay. GSH concentrations were determined by interpolation from a GSH standard curve generated using the bioluminescent system. Exposure to 5mM acetaminophen (APAP) for three hours substantially reduced GSH only after treatment for two days with a P450 inducer, 30 μM pregnenolone 16a-carbonitrile (PCN), whereas exposure to APAP or PCN alone caused little difference in GSH levels.

プロトコール

Complete Protocol

GSH-Glo™ Glutathione Assay Technical BulletinPDF (1238 KB) – English

選択製品の構成品内容

Item	Part #	Size
Reconstitution Buffer with Esterase	V144A	1 × 10ml
Luciferin-NT	V687A	1 × 100μl
GSH-Glo™ Reaction Buffer	V688A	1 × 10ml
Glutathione S-Transferase	V689A	1 × 100μl
Glutathione, 5mM	V690A	1 × 100μl
Luciferin Detection Reagent	V859A	1 × 1 each

SDS

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分析証明書

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使用制限

For Research Use Only. Not for Use in Diagnostic Procedures.

保存条件

選択製品の構成品内容

Item	Part #	Size
Reconstitution Buffer with Esterase	V144B	1 × 50ml
Luciferin-NT	V687B	1 × 500μl
GSH-Glo™ Reaction Buffer	V688B	1 × 50ml
Glutathione S-Transferase	V689B	1 × 500μl
Glutathione, 5mM	V690A	1 × 100μl
Luciferin Detection Reagent	V859B	1 × 1 each

SDS

Search for SDS

分析証明書

Search by lot number

使用制限

For Research Use Only. Not for Use in Diagnostic Procedures.

保存条件

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サイテーション

APD-containing cyclolipodepsipeptides target mitochondrial function in hypoxic cancer cells.

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2018 Cell Chemical Biology
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GSH-Glo™ Glutathione Assay

Part Numbers: V6911, V6912

グルタチオン (GSH) を検出、定量するための発光アッセイ

Want to measure the ratio of reduced to oxidized glutathione?