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|4360S||1 Kit (96 assays)||
|Product Includes||Quantity (with Count)||Solution Color|
|cGMP Rabbit mAb Coated Microwells||1 x 96 tests|
|cGMP-HRP Conjugate||1 x 11 ml||Red|
|cGMP Standard (5 uM)||1 x 1 ml|
|TMB Substrate 7004||1 x 11 ml|
|STOP Solution 7002||1 x 11 ml|
|Sealing Tape||1 x 2 ea|
|ELISA Wash Buffer (20X) 9801||1 x 10 ml|
|Cell Lysis Buffer (10X) 9803||1 x 15 ml|
NOTE: This procedure is for a 96-well tissue culture plate. It can be modified for other tissue culture plates (6, 12, 24, or 48 well).
NOTE: If cell debris is observed it can be removed by brief centrifugation of the plate and transfer of the clear lysates to a new 96 well plate.
NOTE: Watch the color as it being developed since it may be necessary to stop the reaction before 30 minutes.
NOTE: To determine the absolute amount of the tested substance, a standard curve needs to be generated each time. Please follow the detailed protocol on the product datasheet to determine the concentration range for the standard curve.
posted April 2010
Protocol Id: 68
All Species Expected
Cyclic guanosine 3’,5’-monophosphate (cGMP) is a critical and multifunctional second messenger molecule involved in many signal transduction pathways in different cell types of almost all species (1). Intracellular cGMP is generated from GTP by guanylyl cyclase (GC) and degraded through phosphodiesterase (PDE) hydrolysis (1,2). Two distinctive families of GC have been identified: soluble guanylyl cyclases (sGC) that are nitric oxide-responsive and cell membrane-bound, particulate guanylyl cyclases (pGC) that respond to diverse extracellular agonists including peptide hormones, bacterial toxins and free radicals (2,3). Phosphodiesterases form a superfamily of 11 isoforms with different specificity to both cyclic adenosine 3’,5’-monophosphate (cAMP) and cGMP (4). Cyclic GMP regulates cellular physiology by activating cGMP-dependent kinase, modulating cGMP-dependent ion channels or transporters, and altering its own hydrolytic degradation by phosphodiesterase (PDE) (1,4). Because of the diversity of its effectors, cGMP plays an important role in regulating various pathological and physiological processes, such as vascular smooth muscle motility, intestinal fluid and electrolyte homeostasis, and retinal phototransduction (1,5).
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