GSH-Glo™ Glutathione Assay
A Luminescent-Based Assay for the Detection and Quantification of Glutathione (GSH)
- Measure reduced (GSH) or total (GSSG + GSH) glutathione from cells and tissues
- Measure GSH levels directly from cell culture wells
- Excellent sensitivity and easily scalable
Catalog Number:
Size
Catalog Number: V6911
Catalog Number: V6912
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
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
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
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.
Protocols
Complete Protocol
Specifications
Catalog Number:
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 |
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 |
Related Products
類似製品
GSH/GSSG-Glo™ Assay
培養細胞中の総グルタチオン(GSH +GSSG)、GSSG または GSH/GSSG 比を測定するための発光アッセイ
V6611, V6612
Mitochondrial ToxGlo™ Assay
生体異物への暴露によるミトコンドリア機能障害を予測するためのアッセイ
G8000, G8001
ROS-Glo™ H2O2 Assay
培養細胞や酵素反応中の過酸化水素レベルを直接測定できる高感度な発光アッセイ
G8820, G8821
CytoTox-Glo™ Cytotoxicity Assay
細胞集団中の死細胞数を発光で測定する細胞毒性アッセイ
G9290, G9291, G9292
