Product Description
cGMP and cAMP are small cyclic nucleotides that act as intracellular second messengers. cGMP is synthesized by guanylyl cyclase and degraded by phosphodiesterase, playing a critical role in physiological and pathological processes, including cardiovascular function and the regulation of visual and olfactory systems.
The cGMP ELISA Kit uses a competitive enzyme-linked immunoassay (ELISA) method to measure cGMP levels in biological samples. The assay involves competition between HRP-conjugated cGMP and sample cGMP for binding to anti-cGMP monoclonal antibodies. Absorbance at 450 nm is inversely proportional to cGMP concentration. The kit features high sensitivity with an IC50 of ~10 pmol/mL and a detection limit of ~0.1 pmol/mL.
Experimental Overview
- Catalog No: ATK00050
- Detection wavelength: 450 nm (reference 630 nm)
- Sensitivity: 0.1 pmol/mL
- IC50: ~10 pmol/mL
Components & Storage
| Component | Details | Storage |
|---|---|---|
| cGMP Standard | 10000 pmol/mL, 1x0.1mL | -80°C |
| cGMP-HRP | 1000x HRP conjugated with cGMP, 1x0.02mL | -80°C |
| cGMP Antibody | Anti-cGMP monoclonal antibody (1000x), 1x0.02mL | -80°C |
| 10x Assay Buffer | Phosphate buffer with preservative, 1x30mL | 2-8°C |
| Neutralizing Reagent | Tris buffered saline with preservative, 1x8mL | 2-8°C |
| Sample Diluent | 0.1M HCl, 1x30mL | 2-8°C |
| TMB Substrate | 3,3’,5,5’-Tetramethylbenzidine, 1x18mL | 2-8°C |
| Stop Solution | 2M sulfuric acid, 1x8mL | 2-8°C |
| Coated Plate | Goat anti-Mouse IgG, 96 wells | 2-8°C |
Procedure Summary
- Prepare standard solutions, samples, and reagents according to the kit instructions.
- Add Neutralizing Buffer, HCl, standards, or samples to the appropriate wells.
- Add cGMP-HRP conjugate and anti-cGMP antibody to each well (excluding controls).
- Incubate at room temperature for 2 hours.
- Wash the plate 4 times with Assay Buffer and remove residual liquid.
- Add TMB Substrate to each well and incubate for 10 minutes.
- Add Stop Solution and measure absorbance at 450 nm.
Storage & Notes
- Store all reagents as instructed in the kit manual.
- Bring all reagents to room temperature before use.
- Avoid cross-contamination by using clean pipette tips and washing thoroughly.
- Handle Stop Solution with care as it is corrosive.
