Cyclic AMP and GMP XP® Assay Kits
CST社は、Cyclic AMP とCyclic GMP XP® Assay Kits（#4339と#4360）を発売いたしました。競合型イムノアッセイの原理を用いた本キットは、多くのGタンパク質共役受容体（GPCR）の活性測定にご使用いただけます。同キットでは、アッセイプレートに固相化したサイクリックヌクレオチド特異的XP®ラビットモノクローナル抗体に対し、試料中のサイクリックヌクレオチドとキットに含まれる一定量のHPR標識サイクリックヌクレオチドとが競合的に結合します。これにより、吸光度の大きさは試料中のサイクリックヌクレオチドの量に逆比例します。
- 最高品質のXP®モノクローナル抗体を用いることで、高感度かつ高特異性を実現しています。
- 技術サポートは、製品を開発、製造している研究者が担当いたします。
Cyclic GMP XP® Assay Kit #4360
Cyclic Nucleotide EIA Kits
| No. | Name |
|---|---|
| 4339 | Cyclic AMP XP® Assay Kit |
| 4360 | Cyclic GMP XP® Assay Kit |
Figure 1. Cell-based assay: Treatment of RFL-6 cells with sodium nitroprusside (SNP) increases cGMP concentration as detected by Cyclic GMP XP® Assay Kit #4360. RFL-6 cells were seeded at 2x105 cells/well in a 12-well plate and incubated overnight. Cells were either left untreated or pretreated with 0.5 mM IBMX for 15 minutes prior to SNP treatment (30 minutes) and lysed with 1X Cell Lysis Buffer #9803. The absorbance values (left) and percentage of activity (right) are shown above. The percentage of activity is calculated as follows: % activity=100x[(A-Abasal)/(Amax-Abasal)], where A is the sample absorbance, Amax is the absorbance at maximum stimulation (i.e., high SNP concentration), and Abasal is the absorbance at basal level (no SNP). SNP is a nitric oxide donor that directly activates soluble guanylyl cyclases and increases cellular cGMP concentration. IBMX is a non-specific inhibitor of cAMP and cGMP phosphodiesterases that promotes accumulation of cAMP and cGMP in cells.
Cyclic AMP XP® Assay Kit #4339
Figure 2. Cell-based assay: Treatment of 293 cells with isoproterenol increases the cAMP concentration as detected by Cyclic AMP XP® Assay Kit #4339. 293 cells were seeded at 3x104 cells/well in a 96-well plate and incubated overnight. Cells were pretreated with 0.5 mM IBMX for 30 minutes prior to isoproterenol treatment (3 minutes) and lysed with 1X Cell Lysis Buffer #9803. The absorbance values (left) and percentage of activity (right) are shown above. The percentage of activity is calculated as follow: % activity=100*[(A-Abasal)/(Amax-Abasal)], where A is the absorbance of the sample, Amax is the absorbance at maximum stimulation (i.e., high isoproterenol concentration), and Abasal is the absorbance at basal level (no isoproterenol). Isopropterenol is a β-adrenoceptor agonist and activates beta-2 adrenergic receptors (ADRB2) that are endogenously expressed on 293 cells. Activation of ADRB2 then leads to activation of adenylyl cyclase and synthesis of cAMP as its second messenger.