预期应用

ELISA法定量测定大鼠血清、血浆或其它相关液体中环磷酸鸟苷（cGMP）含量。

 

实验原理

本试剂盒应用双抗体夹心酶标免疫分析法测定标本中cGMP水平。用纯化的抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入cGMP抗原、生物素化的抗大鼠cGMP抗体、HRP标记的亲和素，经过彻底洗涤后用底物TMB显色。TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的cGMP呈正相关。用酶标仪在450nm波长下测定吸光度（OD值），计算样品浓度。

 

Intended use

This immunoassay kit allows for the specific measurement of Rat cyclic GMP (cGMP) concentrations in cell culture supernates, serum and plasma.

 

Introduction

Guanosine 3’,5’-cyclic monophosphate (cGMP) is a multi-functional second messenger molecule. It is generated via two pathways distinguished by the types of Guanylyl Cyclase (GC) that mediate its conversion from guanosine triphosphate (GTP). In the soluble pathway, cGMP is generated via cytoplasmic nitric oxide (NO)-activated GC. GCs in the particulate pathway share some homology with those activated by NO but are transmembrane proteins with extracellular ligand-binding domains. The ligands for a subset of membrane GCs are members of the Natriuretic Peptide (NP) hormone family including Atrial NP hormone,B-type NP hormone, and C-type NP hormone.

cGMP primarily affects cellular activities through four different pathways. These include cGMP-dependent Protein Kinases (PKG/GK), cyclic nucleotide-gated (CNG) channels, cAMP-dependent Protein Kinase (PKA), and Phosphodiesterases.PKGs (PKG I and PKG II) are serine/threonine kinases activated by cGMP.

 

Test principle

This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for cGMP has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any cGMP present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for cGMP is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of cGMP bound in the initial step. The color development is stopped and the intensity of the color is measured.