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.
|Sample type||Urine, Serum, Plasma, Other biological fluids, Tissue Extracts, Cell Lysate, Cell culture media|
|Sensitivity||> 0.008 µM|
|Range||0.008 µM – 2 µM|
|Assay time||3h 00m|
COMPONENT AND STORAGE
|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|
|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
Pipettes and pipette tips
Colorimetric microplate reader
96 well plate
100tests ; 200tests ; 500tests
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.