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Competitive ELISA of cGMP using anti-cGMP (RM467). The 96-well plate was coated with 1 ug/mL of Goat anti-rabbit IgG (50 μl/well). 0.05 μg/ml of anti-cGMP (RM467) (50 μl/well) was added and incubated. After wash and block, different concentrations of cGMP (25 μl/well) were added along with 25 μl/well of 1/5,000 diluted HRP conjugated cGMP (71-1359-00). TMB was used to develop the color after incubation and wash.
Competitive ELISA showing the specificity of anti-cGMP (RM467). The 96-well plate was coated with 1 ug/mL of Goat anti-rabbit IgG (50 μl/well). 0.05 μg/ml of anti-cGMP (RM467)(50 μl/well) was added and incubated. After wash and block, cGMP and other cyclic nucleotides or nucleoside phosphates (25 μl/well) were added along with 25 μl/well of 1/5,000 diluted HRP conjugated cGMP (71-1359-00). TMB was used to develop the color after incubation and wash.
Competitive ELISA detecting cGMP in HeLa cells. The 96-well plate was coated with 1 ug/mL of Goat anti-rabbit IgG (50 μl/well). 0.05 μg/ml of anti-cGMP (RM467) (50 μl/well) was added and incubated. After wash and block, HeLa cell lysate (~105cell/mL) or control (lysate buffer only) (25 μl/well) were added along with 25 μl/well of 1/5,000 diluted HRP conjugated cGMP (71-1359-00). TMB was used to develop the color after incubation and wash.
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