Figure 1: cGMP Standard was diluted in 1X Cell Lysis Buffer #9803 and samples were assayed following the Cyclic GMP XP ® Assay Kit protocol. This standard curve is for demonstration purposes only; users should generate a standard curve for each sample set in order to accurately determine cGMP concentration.
![Figure 2: 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 2x10 5 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-A basal )/(A max -A basal )], where A is the sample absorbance, A max is the absorbance at maximum stimulation (i.e., high SNP concentration), and A basal 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.](http://www.bioprodhub.com/system/product_images/ak_products/1/sub_1/1757_4360_fig2_jp.jpg)
Figure 2: 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 2x10 5 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-A basal )/(A max -A basal )], where A is the sample absorbance, A max is the absorbance at maximum stimulation (i.e., high SNP concentration), and A basal 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.
![Figure 3: Treatment of RFL-6 cells with atrial natriuretic peptide (ANP) increases cGMP concentration as detected by Cyclic GMP XP ® Assay Kit #4360. RFL-6 cells were seeded at 2x10 5 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 ANP 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-A basal )/(A max -A basal )], where A is the sample absorbance, A max is the absorbance at maximum stimulation (i.e., high ANP concentration), and A basal is the absorbance at basal level (no ANP). ANP is a ligand that binds and activates membrane bound guanylyl cyclases to increase cellular cGMP concentration.](http://www.bioprodhub.com/system/product_images/ak_products/1/sub_1/1758_4360_fig3_jp.jpg)
Figure 3: Treatment of RFL-6 cells with atrial natriuretic peptide (ANP) increases cGMP concentration as detected by Cyclic GMP XP ® Assay Kit #4360. RFL-6 cells were seeded at 2x10 5 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 ANP 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-A basal )/(A max -A basal )], where A is the sample absorbance, A max is the absorbance at maximum stimulation (i.e., high ANP concentration), and A basal is the absorbance at basal level (no ANP). ANP is a ligand that binds and activates membrane bound guanylyl cyclases to increase cellular cGMP concentration.