cAMP-Glo™ Max Assay
Faster cAMP Detection for High-Throughput Assays
- Complete in ~30 minutes
- Scalable to 1536-well plate formats and beyond
- No interference by fluorescent compounds
Catalog Number:
Size
Catalog Number: V1681
Catalog Number: V1682
Catalog Number: V1683
Bioluminescent Detection of cAMP to Monitor GPCR Activity
cAMP-Glo™ Max Assayは96、384または1536ウェルプレートで可能です。実験では、cAMPレベルが変化する適切な時間をかけて細胞をテスト化合物で誘導します。誘導後に細胞を溶解し、放出されたcAMPが試薬に含まれるprotein kinase Aを刺激します（Figure 1）。次にKinase-Glo® Reagentを加えてPKA反応を停止し、残存するATPをルシフェラーゼ反応で検出します。プレートの測定はマイクロプレートルミノメーターで行います。発光シグナルの半減期は4時間以上です。シグナルの半減期が長いため、ルミノメーターにインジェクターは不要で、マルチプレートのバッチモード処理を可能にします。
- 迅速で簡便：細胞溶解およびcAMP検出が1ステップに統合（cAMP-Glo™ ONE Buffer）、アッセイは約30分で完了
- 優れたシグナル/バックグラウンド比（S/B比） ：優れたS/B比（>200 ［cAMP］，>15 ［細胞］）を示し、1536ウェルプレート以上のフォーマットにも簡単に変更可能
- 柔軟性：ONE Buffer：5X濃度は細胞培養ボリュームの柔軟性を向上
- 優れた発光技術を採用：定評のあるUltra-Glo™ Recombinant Luciferaseを使用、蛍光化合物による干渉も無し、Non-RI
- GPCR活性に対するモジュレーターを同定するための化合物ライブラリーを用いたハイスループット分析
- 様々な細胞種で簡単にcAMP定量が可能
【パフォーマンス】製品概説（原理、アゴニスト、アンタゴニストタイトレーション、組織抽出液での使用例）
カタログ番号V1681は96または384プレート2枚分のアッセイに十分な量の試薬です。
カタログ番号V1682は96または384プレート20枚分のアッセイに十分な量の試薬です。
カタログ番号V1683は96または384プレート200枚分のアッセイに十分な量の試薬です。
Improved Format for Better Performance
This improved version of the cAMP-Glo™ Assay combines the lysis and cAMP reaction buffers into the cAMP-Glo™ ONE Buffer, streamlining the protocol and speeding up the assay. The ONE Buffer is supplied at 5X concentration, which provides increased flexibility for starting cell culture volumes.
The cAMP-Glo™ Max Assay allows easy quantitation of cAMP levels in broad range of cell types, and can be performed in 96-, 384- or 1536-well plates. Cells are induced with a test compound for an appropriate period of time to modulate cAMP levels. After induction, cells are lysed and the cAMP released stimulates protein kinase A in the reagent. The Kinase-Glo® Reagent is then added to terminate the PKA reaction and detect the remaining ATP via a luciferase reaction. Plates are read using a microplate-reading luminometer. The half-life for the luminescent signal is greater than 4 hours, providing ample time to read plates and eliminating the need for luminometers with reagent injectors.
Cat.# V1681 contains sufficient reagents for 2 plates (96- or 384-well format).
Cat.# V1682 contains sufficient reagents for 20 plates (96- or 384-well format).
Cat.# V1683 contains sufficient reagents for 200 plates (96- or 384-well format).
cAMP production in cells and the cAMP-Glo™ Max Assay.
Schematic diagram of cAMP production in cells and the cAMP-Glo™ Max Assay.
Fast and Easy to Use
- Improved—Lysis and cAMP detection steps combined (cAMP-Glo™ ONE Buffer).
- ONE Buffer—5X concentration provides better flexibility for starting cell culture volumes.
- Assay complete in approximately 30 minutes.
Excellent Signal:Noise Ratios
- Best signal:background ratio of all the cAMP assays.
- Signal:Background >200 (with cAMP), >15 (on cells).
Proven Luminescence Technology
- Powered by Ultra-Glo™ Recombinant Luciferase.
- No interference by fluorescent compounds.
- Non-radioactive.
Protocols
Complete Protocol
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
|
Protein Kinase A |
V135A | 1 × 20μl |
|
cAMP-Glo™ ONE Buffer |
V166A | 1 × 2.5ml |
|
MgCl2, 1M |
V330A | 1 × 1ml |
|
cAMP, 1mM |
V642B | 1 × 100μl |
|
Kinase-Glo® Substrate |
V672A | 1 × 1 vial |
|
Kinase-Glo® Buffer |
V673A | 1 × 10ml |
U.S. Pat. No. 7,700,310 and other patents and patents pending.
選択製品の構成品内容
| Item | Part # | Size | Concentration |
|---|---|---|---|
|
Protein Kinase A |
V135B | 1 × 200μl | |
|
cAMP-Glo™ ONE Buffer |
V166B | 1 × 20ml | |
|
MgCl2 |
V330B | 1 × 5ml | 1M |
|
cAMP, 1mM |
V642A | 1 × 500μl | |
|
Kinase-Glo® Substrate |
V672B | 1 × 1 vial | |
|
Kinase-Glo® Buffer |
V673B | 1 × 100ml |
U.S. Pat. No. 7,700,310 and other patents and patents pending.
選択製品の構成品内容
| Item | Part # | Size | Concentration |
|---|---|---|---|
|
Protein Kinase A |
V135B | 10 × 200μl | |
|
cAMP-Glo™ ONE Buffer |
V166B | 10 × 20ml | |
|
MgCl2 |
V330B | 4 × 5ml | 1M |
|
cAMP, 1mM |
V642A | 6 × 500μl | |
|
Kinase-Glo® Substrate |
V672B | 10 × 1 vial | |
|
Kinase-Glo® Buffer |
V673B | 10 × 100ml |
U.S. Pat. No. 7,700,310 and other patents and patents pending.
Resources
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