DESCRIPTION
cGMP Assay Kit ab65356 provides a direct competitive immunoassay for sensitive and quantitative determination of cGMP level in biological samples.The cGMP assay protocol uses the recombinant Protein G coated plate to anchor cGMP polyclonal antibody. cGMP-HRP conjugates directly competes with cGMP from samples for binding to the cGMP specific antibody on the plate. After incubation and washing, the amount of cGMP-HRP bound to plate can easily be determined by reading OD450nm. The intensity of OD450nm is inversely proportional to the concentration of cGMP in samples.The kit provides a new acetylation procedure that improves detection signal significantly. The kit can detect 0.04-10 pmol/well (0.008-2 µM) cGMP samples.
Detection method | Colorimetric |
Sample type | Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media |
Assay type | Quantitative |
Sensitivity | > 0.008 µM |
Range | 0.008 µM – 2 µM |
Assay time | 3h 00m |
COMPONENT AND STORAGE
A.Kit Components
Item | Quantify | Storage Temp |
10X cGMP Assay Buffer | 25 mL | +4°C |
Standard cGMP (10 nmol) | 1 vial | -20°C |
Neutralizing Buffer | 7.5 mL | +4°C |
Acetylating Reagent A | 0.75 mL | +4°C |
Acetylating Reagent B** | 1.5 mL | +4°C |
Anti-cGMP pAb/BSA | 1 vial | -20°C |
cGMP-HRP/BSA | 1 vial | -20°C |
HRP Developer | 10 mL | +4°C |
Protein G Coated Plate | 1 each | -20°C |
* Store kit at -20°C.
Dilute the 10X cGMP Assay Buffer to 1X Assay Buffer with ultra-pure water. Store at 4°C.
Reconstitute the Standard cGMP (pellet may not be visible) in 1 ml of 0.1M HCl (not provided), vertex for 10 seconds to generate 10 pmol/μl cGMP stock standard solution.
Reconstitute rabbit anti-cGMP pAb and cGMP-HRP each with 1.1 ml of the 1X Assay Buffer as stock solutions.
Unused well strips can be kept at –20°C with the desiccants, stable for up to 1 month.
The kit should be stored at –20°C. After reconstitution, some components may be stored at 4°C as instructed above, stable for up to 1- 2 months.
*NOTE: Acetylating Reagent B is very volatile and hence the vial has to be tightly capped and stored only at +4°C.
B. Additional Materials Required
Microcentrifuge
Pipettes and pipette tips
Colorimetric microplate reader
96 well plate
Orbital shaker
HCl
SIZE
100tests ; 200tests ; 500tests
USAGE
Celltechgen provides high-quality reagents and materials for research use only. For proper handling of potentially hazardous chemicals, please request the Safety Data Sheet (SDS) provided for the product.
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